EUROIMMUN

EUROIMMUN
Medizinische Labordiagnostika AG
Product Catalogue
Diagnostics for the Determination of Autoantibodies, for Infectious Serology and Allergology Indirect Immunofluorescence ELISA RIA Westernblot EUROASSAY EUROLINE EUROPLUS
2010
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EUROIMMUN
Table of Contents
EUROIMMUN Company Profile .................................................................................................................................................. 3 Techniques for the Serological Investigation of Antibodies ....................................................................................................... 5 Indirect Immunofluorescence: An Easy and Modern Method ............................................................................................................................... 6 BIOCHIP Mosaics ..................................................................................................................................................................................................... 9 The Indirect Immunofluorescence Test, Performed Using the TITERPLANE Technique .............................................................................. 10 Recommended Serum Dilutions for Indirect Immunofluorescence ................................................................................................................... 12 Diagnostically Relevant Systemic Autoantibodies ............................................................................................................................................... 16 Organ-/Tissue-Specific Autoantibodies .................................................................................................................................................................. 17 Antibodies for Infectious Serology ......................................................................................................................................................................... 18 Antibodies for Allergology ....................................................................................................................................................................................... 19 EUROIMMUN Microplate ELISA .............................................................................................................................................................................. 18 ELISA Automation using the EUROIMMUN Analyzer I ........................................................................................................................................ 22 Incubating the Microplate ELISA ............................................................................................................................................................................. 23 EUROASSAY: Line Blots in TITERPLANETechnique Format ........................................................................................................................... 24 Incubating the EUROASSAY (TITERPLANETechnique) .................................................................................................................................... 25 The EUROLINE: A New Technique for Extensive Antibody Profiles .................................................................................................................. 26 EUROLINE Automation Using EUROBlotMaster and EUROLineScan ................................................................................................................ 27 Westernblots/EUROLINE-WB: Reliable Differentiation of Antibodies Present .................................................................................................. 28 Incubating the EUROLINE/Westernblot/EUROLINE-WB ....................................................................................................................................... 29 EUROIMMUN Radioimmunoassays (RIA/IRMA) ................................................................................................................................................... 30 EUROIMMUN Products for the Determination of Autoantibodies ............................................................................................ 31 Autoantibodies against Cell Nuclei (ANA) ............................................................................................................................................................. 32 Autoantibodies against Double-Stranded DNA (dsDNA) ..................................................................................................................................... 35 Autoantibodies against CCP and Sa ....................................................................................................................................................................... 36 Autoantibodies against Mitochondria (AMA) ........................................................................................................................................................ 37 Autoantibodies against Liver Antigens .................................................................................................................................................................. 38 Autoantibodies against Thyroid Gland Antigens / Antigen Detections ............................................................................................................. 40 Autoantikrper against Antigens of the Skin ........................................................................................................................................................ 41 Autoantibodies against Neuronal Antigens ........................................................................................................................................................... 42 Autoantibodies against Islet Cell Antigens ............................................................................................................................................................ 44 Autoantibodies against Parietal Cells (PCA) .......................................................................................................................................................... 45 Autoantibodies against Granulocyte Cytoplasm (cANCA/pANCA) ..................................................................................................................... 46 Antibodies against Endomysium and Gliadin ....................................................................................................................................................... 48 EUROIMMUN Products for Infectious Serology ........................................................................................................................ 49 Antibodies against Borrelia ...................................................................................................................................................................................... 50 Antibodies against Epstein-Barr Virus (EBV) ......................................................................................................................................................... 52 Antibodies against Helicobacter Pylori .................................................................................................................................................................. 54 Antibodies against Herpes Simplex Virus (HSV) .................................................................................................................................................. 55 Antibodies against Chlamydia ................................................................................................................................................................................. 56 Antibodies against Emerging Viruses .................................................................................................................................................................... 57 BIOCHIP Mosaics for Infectious Serology .......................................................................................................................................................... 58 Additional Reagents for the Determination of Acute Infections ......................................................................................................................... 60 EUROIMMUN Products for Allergology ..................................................................................................................................... 61 Order Information and Product Data ......................................................................................................................................... 64 Fluorescence-Labelled Antibodies: Fluorescein (FITC) for EUROIMMUN IIFT .................................................................................................. 65 Controls for EUROIMMUN IIFT: Organ-Specific Autoantibodies ........................................................................................................................ 66 Controls for EUROIMMUN IIFT: Systemic Autoantibodies .................................................................................................................................. 68 Controls for EUROIMMUN IIFT: Infectious Serology ............................................................................................................................................ 70 Controls for EUROIMMUN IIFT: Determination of Further Antibodies .............................................................................................................. 76 Controls for EUROIMMUN Westernblots/EUROLINE-WB .................................................................................................................................... 77 EUROASSAY for the Determination of Autoantibodies (Test Systems) ............................................................................................................ 79 EUROLINE for the Determination of Autoantibodies (Test Systems) ................................................................................................................ 80 EUROLINE for Infectious Serology (Test Systems) .............................................................................................................................................. 81 EUROLINE for Allergology (Test Systems) ............................................................................................................................................................ 81 Westernblot/EUROLINE-WB for the Determination of Autoantibodies (Test Systems) ................................................................................... 83 Westernblot/EUROLINE-WB for Infectious Serology (Test Systems) ................................................................................................................. 84 Microplate ELISA for the Determination of Autoantibodies (Test Systems) ..................................................................................................... 86 Latex Agglutination tests for the Determination of Autoantibodies (Test Systems) ....................................................................................... 89 EUROArray for Molecular Genetic Determinations (Test Systems) ................................................................................................................... 89 Radioimmunoassay (RIA) for the Determination of Autoantibodies / Autoantigens / Hormone Determination (Test Systems) ............... 89 Microplate ELISA for Infectious Serology (Test Systems) ................................................................................................................................... 91 Microplate ELISA for the Determination of Antibodies against Other Antigens (Test Systems) ................................................................... 95 Allercoat 6 System ................................................................................................................................................................................................ 96 Diagnostics for Indirect Immunofluorescence: Organ-Specific Autoantibodies ............................................................................................. 116 Diagnostics for Indirect Immunofluorescence: Systemic Autoantibodies ....................................................................................................... 126 Diagnostics for Indirect Immunofluorescence: Infectious Serology ................................................................................................................. 134 Diagnostics for Indirect Immunofluorescence: Other Antigens ........................................................................................................................ 151 Further Reagents for EUROIMMUN IIFT .............................................................................................................................................................. 151 Other Items for EUROIMMUN IIFT ........................................................................................................................................................................ 152 General Delivery Conditions .................................................................................................................................................................................. 153 Index ......................................................................................................................................................................................... 154
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EUROIMMUN
EUROIMMUN COMPANY PROFILE
Company site Gro Grnau
Company headquarters Lbeck
Company branch Dassow
Company branch Rennersdorf
Company branch Pegnitz
EUROIMMUN was founded in September 1987 and today has its headquarters in Lbeck, Germany. Branches are situated in Gro Grnau near Lbeck (Schleswig-Holstein), in Rennersdorf (Upper Lusatia, Saxony), Dassow (Mecklenburg-Western Pomerania) and in Pegnitz (Upper Franconia, Bavaria). Further EUROIMMUN subsidiaries can be found in Canada (Mississauga), China (Beijing, Hangzhou), Great Britain (Pontypool in Wales), Italy (Padua), Lebanon (Beirut), Poland (Wroclaw), Switzerland (Lucerne), Singapore, South Africa (Capetown), Turkey (Istanbul) and the USA (New Jersey). At present EUROIMMUN has 714 employees in Germany, 884 worldwide. The company is ISO-certified (EN ISO 9001:2008, EN ISO 13485:2003/ CMDCAS). EUROIMMUN produces reagents for medical laboratory diagnostics. In the foreground are test systems for the determination of various antibodies in patient serum in the diagnosis of autoimmune diseases, infectious diseases and allergies. The test methods employed are predominantly indirect immunofluorescence, microplate ELISA, various blot techniques (Westernblot, EUROASSAY, EUROLINE, EUROLINE-WB) and all molecular biology techniques. The company is based on worldwide-patented state-of-the-art production methods and microanalysis techniques and is one of the worlds leading manufacturers of medical laboratory diagnostics. The BIOCHIPs are one of EUROIMMUNs many inventions: paper-thin sheets of glass are coated with cells or tissue sections and then cut automatically into millimetre-sized fragments which are subsequently glued onto slides using a fully automated device. This BIOCHIP technology allows extreme miniaturization and standardization of immunbiochemical analyses. With BIOCHIP Mosaicsmade from 30 or more different organ sections, cell substrates or defined antigens (EUROPLUS) only minimum incubation efforts are necessary to obtain a detailed antibody profile. In EUROIMMUN enzyme immunoassays (ELISA) defined antigens, purified using state-of-the-art biotechnological processes, are employed as the antigen substrate. Some of these antigens are synthesized in the companys molecular biology laboratories. EUROIMMUN ELISA are characterized by their excellent stability, simple handling and short incubation times, and they are ideal for automated use. All reagents are delivered ready-to-use and are exchangeable between different lots. EUROIMMUN offers the largest and most differentiated arsenal of enzyme immunoassays worldwide for the diagnosis of autoimmune and infectious diseases.
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EUROIMMUN
The innovative procedures EUROASSAY and EUROLINE developed by EUROIMMUN follow the same test principle as the ELISA methods, with the use of the BIOCHIP technology. At EUROIMMUN purified antigens are printed in parallel lines at defined positions on membrane strips. Following incubation, users can evaluate results visually without additional equipment. These reagents allow in particular the differentiation of antibodies that are not clearly defined microscopically by indirect immunofluorescence. EUROASSAY and EUROLINE are also employed in laboratories where no sophisticated laboratory instruments are available. EUROIMMUN produces an extensive range of Westernblot strip test systems and corresponding reagents for confirmation of positive fluorescence and ELISA results as well as clarification of difficult-to-interpret results in autoimmune diagnostics, infectious serology and allergology. Refined electrophoretical processes have been developed to allow the precise separation of diagnostically relevant proteins from one another. Lot-specific evaluation templates are produced for the evaluation of band patterns. The program EUROLineScan enables fully automated evaluation of membrane-based test systems and simplifies the archiving of results with large sample series. One of the companys main strengths is its technical expertise. This encompasses not just the manufacture and sale of medical laboratory diagnostics, but also the diagnostic application of the products in a reference laboratory which provides highly differential diagnostics. This reference laboratory has set standards in Germany, and worldwide is unequalled in the whole field of autoimmune diagnostics. The diagnostic spectrum of the laboratory also covers the areas of infectious serology and serological allergy diagnostics. The reference laboratory receives hundreds of serum samples daily from all over Germany as well as from many other countries. It helps EUROIMMUN customers to secure their results: a large proportion of serum samples sent to EUROIMMUN for evaluation are analysed free of charge in order to maintain high standards in the laboratories of EUROIMMUN customers. Customers can obtain further technical information from experienced scientists in the company, with whom they can also discuss serological problem cases. The Institute for Quality Assurance, an institution newly founded by EUROIMMUN, organises unbiased quality assessments and provides advice in the area of quality management. Moreover EUROIMMUN has established the Institute for experimental Immunology, which is engaged in basic research. In October 2009, the EUROIMMUN workforce included 134 university and college graduates, among these biologists, biochemists, chemists, engineers and medical doctors (40 of them holding a doctors degree). Medical technicians are particularly strongly represented with 116 people, corresponding to EUROIMMUNs activities, as well as biology/chemistry laboratory technicians (55). At present the company is training 51 young people as biology laboratory assistants, industrial clerks, IT specialists, electronic system technicians, electronic technicians for devices and systems, industrial mechanics, lathe operators and cooks as well as business information technology specialists and business economists (dual system). At EUROIMMUN great value is placed on advising customers and prospective customers in a factual, technical and commercially restrained manner and fully supporting them in the use of our diagnostically demanding products. EUROIMMUN products are backed by an energetic and competent sales force, qualified information material, didactic test instructions and scientifically based, but nevertheless understandable advertisements in technical journals. Advertising material is produced in-house using the latest desktop publishing methods, right up until the fully digitalised ready-for-exposure documents. The most important publications and posters are translated into many languages. EUROIMMUN has set up an informative homepage on the internet (www.euroimmun.com) which is visited extensively internationally. Over 3,000 laboratories worldwide use EUROIMMUN diagnostics. 400 of these are in Germany. The companys development is shaped by continuous growth. Although the diagnostic market in Germany stagnated and has become particularly strongly competitive, EUROIMMUN has been able to continue its strong expansion. The company is achieving an ever increasing independence from the German market, since more and more products are sold abroad. With their quality and standardization, EUROIMMUN products are capturing the leading position in the world.
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EUROIMMUN
TECHNIQUES FOR THE SEROLOGICAL INVESTIGATION OF ANTIBODIES
Techniques for the Serological Investigation of Antibodies
EUROIMMUN
Indirect Immunofluorescence: An Easy and Modern Method

Principle of the Test
cell with antigen
For the determination of autoantibodies or antibodies against infectious agents, cells, tissue sections or purified, biochemically characterized substances are used as antigen substrates. If the sample is positive, specific antibodies in the diluted serum sample attach to the antigens coupled to a solid phase. In a second step, the attached antibodies are stained with fluorescein-labelled anti-human antibodies and visualized with the fluorescence microscope. Positive samples can be titrated in steps. The most suitable titration interval is provided by the dilution factor 3.162 (square root of 10). In this way, every second step represents in its denominator an integral power of 10 (1 : 10, 1 : 32, 1 : 100, 1 : 320, 1 : 1000, 1 : 3200, 1 : 10000 etc.).
specific human antibody
FITC
FITC-labelled antihuman antibody
FITC
Indirect Immunofluorescence: A Standardized Technique for the Determination of Autoantibodies and Antibodies against Infectious Agents
High specificity: positive and negative samples produce a large difference in signal strength. Each bound antibody shows a typical fluorescence pattern depending on the location of the individual antigens. The entire antigen spectrum of the original subtrate is available, thus allowing the detection of a large number of antibodies and achieving a higher detection rate. Immunofluorescence enables simultaneous detection of antibodies against several biochemically different antigens on one single biological substrate. The indirect immunofluorescence test is the analytical method of choice when it would be too difficult or too complicated to prepare the test antigens individually for enzyme immunoassays.
fine-granular. Pattern homog. Anti- Pattern dsDNA? Anti-Histones? Anti-SS-A? Anti-SS-B?
Pattern nucleolar. Anti- Pattern cytoplasmic. PM-Scl? AMA M2? Differentiation of antibodies using HEp-2 cells. tissue sections antigen dots culture cells transf. cells
EUROIMMUNs Innovations for the Standardization and Modernization of Indirect Immunofluorescence

Activation technique: physically or chemically activated cover glasses are coated with cultured cells or tissue sections. Frozen tissue sections are fixed to the glass surface by covalent bonding, increasing adhesion more than 100 times and thus preventing the substrates from being detached. BIOCHIP Technology: cover glasses coated with biological substrates are cut into millimetre-sized fragments (BIOCHIPs) on a machine. This makes it possible to obtain ten or more first-class preparations of homogeneous quality per tissue section, in the case of cultured cell substrates even several thousands. BIOCHIP Mosaics: using several BIOCHIPs coated with different substrates side by side on one and the same reaction field, antibodies against various organs or infectious agents can be investigated simultaneously. Detailed antibody profiles can thus be established with comparatively little effort, allowing the reciprocal determination of the results on different substrates. TITERPLANE Technique: samples or reagents are applied to the reaction fields of a reagent tray. The BIOCHIP Slides are then placed into the recesses of the reagent tray, where all BIOCHIPs come into contact with the fluids, and the individual reactions commence simultaneously. As the fluids are confined in a closed space, there is no need for the use of a conventional humidity chamber.
BIOCHIP Technology and Mosaics.
EUROIMMUN
Chemically Activated Cover Glasses for Histochemistry

For diagnostics of organ-specific or tissue-specific autoantibodies frozen tissue sections of various organs are used. However, formerly, the morphology of tissues suffered during incubation in aqueous medium, tissue parts occasionally became detached from slides, and the interpretation of results was difficult. Using the activation technique for the first time in histology, we have applied solid phase techniques. Firstly, the surface of cover glasses is coated with spontaneously reactive aldehyde groups. In a second step, the tissue sections are applied to the chemically activated cover glasses (Stcker, W: European Patent No. 0 117 262; U.S. Patent No. 4,647,543). Free amino groups of the tissue sections, especially of the hydroxylysine contained in the collagen, bind to the carrier material by covalent bonding. This results in an increased adhesion of frozen tissue sections more than a hundredfold and prevents them from being detached during incubation. Furthermore, in some cases the activation technique results in a significantly better conservation of tissue structures, especially in organs which previously exhibited a generally low level of adhesion. Therefore, the tests can be evaluated with considerably greater confidence.
frozen tissue section
frozen tissue section N
glutardialdehyde
Aminoethylaminoproyltrimethoxysilane NH2
(CH2)2 NH
(CH2)3
- 3 CH3OH
HC
NH2
HC
(CH2)3
HC
O
HC
(CH2)3
(CH2)3
HC
HC
- H 2O
NH2
- H2O
N
(CH2)2 NH
(CH2)3
Si
N
(CH2)2 NH
(CH2)3
Si
(CH2)2 NH
H3CO
Si
OCH3
OCH3
(CH2)3
Si
OH
glass
Fixation of frozen tissue sections to glass surfaces by covalent bonding.
Determination of Low-Avidity Antibodies

An alternative principle for the serological diagnosis of fresh infections has been established by investigating the antibody avidity. The first reaction of the immune system following an infection is the formation of low-avidity antibodies. As the infection proceeds, increasingly antigen-adapted IgG is formed, and avidity grows. As long as high-avidity IgG is not yet detected in the serum, it can be assumed that the infection is still in an early stage. To identify low-avidity antibodies in a patients serum, two immunofluorescence tests are performed in parallel: one test is carried out in the conventional way, the other one includes urea treatment between incubations with patients serum and peroxidase-labelled anti-human IgG, resulting in the detachment of lowavidity antibodies from the antigens. Low-avidity antibodies are present if the fluorescence intensity is significantly reduced (two intensity levels ore more) by urea treatment. The following test kits for avidity determination are available: Toxoplasma gondii, Rubella virus, West-Nile virus, CMV, EBV-EA, EBV-CA.
low-avide Ab against EBV-CA
high-avide Ab against EBV-CA without urea with urea
EUROPLUS System: Combination of conventional immunofluorescence substrates and monospecific tests

In EUROPLUS immunofluorescence tests antibody detection is performed using both tissue sections/cell substrates and monospecifically reacting antigens. Antibodies detected in IFT screening tests can therefore be differentiated or confirmed with one and the same reaction field. In some cases, the antigens help to extend the antigen spectrum, offering a wider range for screening. BIOCHIPs coated with purified or recombinant antigens are used as monospecific substrates. In case of a positive result the antigens fluoresce green in defined areas under the microscope. In some EUROPLUS test systems several different antigens are coated on one BIOCHIP in separate antigen rows. In this manner, several monospecific analyses can be performed using a single BIOCHIP. Available EUROPLUS substrates: MPO, PR3, gliadin GAF-3X, OspC, VlsE, Plasmodium falciparum und vivax, gp125, p19.
EUROPLUS: BIOCHIP combination of tissue sections / cell substrates (left) and purified antigens (right).
EUROIMMUN

New: Hydrophobic Slides
EUROIMMUN has developed a specific slide surface with hydrophobic properties. This prevents the droplets from spreading on the slide surface. The slides are now suitable for manual incubation using TITERPLANE Technique and nonmanual incubation on automated systems. The reaction fields must not be marked with a Cytomation Pen anymore. Price and order number are the same as for conventional EUROIMMUN slides (please add "hydrophobic" when ordering) Hydrophobic slides are available on request for many products.
Above: conventional slide Below: hydrophobic slide.
AP16 IF Plus by DAS: Automated Solution for all EUROIMMUN Immunofluorescence Tests
AP16 IF Plus by DAS. Various validated parameters. Slide definitions and test files available. CE conformity for device/test system combination. Capacity: 16 slides, 80 samples, 200 dilutions. Programmable for 8 methods per run. 12 dilution series freely programmable. Automated sample dilution, sample and reagent dispension, incubation and washing of slides. Laboratory software interface. The incubation protocol for result documentation is automatically created from the worklist. Barcode reader available on request.
Fluorescence Microscope EUROStar II

The EUROStar II is specifically tailored to the requirements of indirect immunofluorescence. Unnecessary and partly expensive components have been deliberately left out of the design and the conventional complex illumination fittings have been replaced by the stunningly simple EUROStar Bluelight system. With the EUROStar Bluelight, engineers at EUROIMMUN AG have introduced blue light-emitting diodes to fluorescence microscopy. Almost all the emitted light is suitable for the excitation of fluorescein. The EUROStar Bluelight does not emit any ultraviolet radiation and is explosion proof. The EUROStar Bluelight is immediately ready for operation after being switched off and offers instant full output after being switched on again. The LED shines for 50,000 hours which is 500 times longer than a mercury vapour lamp. Thus, the microscope requires almost no maintenance. With its halogen transmitted-light source, the version EUROStar II Plus is suited for brightfield, darkfield, phase contrast or polarisation microscopy. EUROStar was designed to accommodate the additional assembly of a digital camera.
EUROIMMUN
Slides for indirect immunofluorescence can be produced with single substrates or BIOCHIP Mosaics from up to 45 different substrates according to your individual requirements. thyroid gland, parathyroid gland, pancreas, adrenal gland, ovary, placenta*, testis, spermatozoa, pituitary gland, hypothalamus*, cerebrum, cerebellum, brain stem, pons*, lobus temporalis, substantia nigra*, peripheral nerve, spinal cord, optic nerve (AQP-4, NMO IgG) eye, granulocytes (fixed with EOH, HCHO or MOH), lactoferrin-specific granulocytes, lymphocytes, monocytes*, thrombocytes, kidney (primate, rat, mouse), lung*, liver (primate, rat, mouse), mouth mucosa, stomach (corpus, antrum), jejunum, colon, intestinal goblet cells, umbilical cord, mamma, lacrimal gland, parotid gland, prostate, vesicula seminalis, skeletal muscle, F-actin (VSM47), heart muscle, thymus, lipocytes, cartilage*, epidermis, oesophagus (primate, rat), tongue, lip, melanocytes, HEp-2 cells, HEp-20-10 cells, HUVEC, Crithidia luciliae sensitive etc. Adenovirus, Afipia felis*, Bartonella henselae, B. quintana, Bordetella parapertussis, B. pertussis, Borrelia afzelii, B. burgdorferi sensu stricto (strains CH, USA), B. garinii, Campylobacter coli*, C. jejuni, Candida albicans, C. glabrata*, C. krusei*, C. parapsilosis*, C. tropicalis*, Chikungunya virus, Chlamydia pneumoniae, C. trachomatis, C. psittaci, CMV, Coxsackievirus (A7, A9, A16, A24, B1 to B6), Crimean Congo fever virus, Dengue virus type 1 to 4, EBV-CA, EBNA, EBVEA, Echinococcus granulosus, ECHO virus, Hantavirus, Haemophilus influenzae*, Helicobacter pylori, HHV-6, HSV-1, HSV-2, Influenza virus A (strains H3N2, H1N1, H5N1), Influenza virus B, Japanese encephalitis virus, Klebsiella pneumoniae*, Legionella bozemanii*, L. dumoffii*, L. gormanii*, L. jordanis*, L. longbeachae, L. micdadei*, L. pneumophila (serotypes 1 to 14), Leishmania donovani, Listeria monocytogenes (1/2a and 4b)*, measles virus, mumps virus, Mycoplasma hominis, M. pneumoniae, Parainfluenza virus type 1 to 4, Rift valley fever virus*, RSV, rubella virus, Saccharomyces cerevisiae, SARS-CoV, TBE virus, TO.R.C.H. profile, Toxoplasma gondii, Treponema pallidum, T. phagedaenis, Ureaplasma urealyticum, VZV, West Nile virus, Yellow fever virus, Yersinia enterocolitica (O:3, O:4, O:6 and O:9)*. EUROPLUS: HEp-2/liver + RNP/Sm, Sm, SS-A, SS-B, Scl-70, rib. P-proteins, Jo-1; granulocytes + MPO, PR3; primate stomach (parietal cells) + intrinsic factor; primate liver (endomysium) + gliadin (GAF-3X); rat kidney + AMA M2; thyroid gland + thyroglobulin; Borrelia burgdorferi and afzelii + OspC and VlsE, Plasmodium falciparum (HRP-2, MSP-2), P. vivax (MSP, CSP). Transfected cells: rPAg 1 + 2 (pancreas antigen 1 + 2), AQP-4, glutamate receptor (type NMDA), desmoglein 1 + 3, BP230. * Currently not available in the European Union.
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BIOCHIP Mosaics
EUROIMMUN
The Indirect Immunofluorescence Test, Performed Using the TITERPLANE Technique

(Reaction fields 5 x 5 mm)
The TITERPLANE Technique was developed by EUROIMMUN in order to standardize immunological analyses: Samples or labelled antibodies are applied to the reaction fields of a reagent tray. The BIOCHIP Slides are then placed into the recesses of the reagent tray, where all BIOCHIPs of the slide come into contact with the fluids, and the individual reactions commence simultaneously. Position and height of the droplets are exactly defined by the geometry of the system. As the fluids are confined in a closed space, there is no need for the use of a conventional humidity chamber. It is possible to incubate any number of samples next to each other and simultaneously under identical conditions. Prepare: Check the reagent tray: Are the reaction fields hydrophiIic and the surrounding coating hydrophobic? If not, rub with a wet paper towel, using normal household detergent or Extran MA 01 (Merck) if necessary, and rinse thoroughly with water. For occasional disinfection, immerse for 1 h in 3% Sekusept Extra (Henkel) in water. Open the individual packets containing the BIOCHIP Slides only after they have reached room temperature. Do not touch the BIOCHIPs. Mark BIOCHIP Slides as required with a felt pen. Dilute: Dilute serum samples according to the users test protocol. Include positive and negative controls with every test procedure. Mix control sera before use. Pipette: Apply 25 l of diluted serum to each reaction field of the reagent tray, avoiding air bubbles. Transfer all samples to be tested before starting the incubation (up to 200 droplets). Use a polystyrene pipetting template. Incubate: Start reactions by fitting the BIOCHIP Slides into the corresponding recesses of the reagent tray. Ensure that each sample makes contact with its BIOCHIP and that the individual samples do not come into contact with each other. Incubate for 30 min at room temperature. Wash: Rinse the BIOCHIP Slides with a flush of PBS-Tween using a beaker, and immerse them immediately afterwards in a cuvette containing PBS-Tween for at least 5 min. Pipette: Apply 20 l of fluorescein-labelled antihuman immunoglobulin (conjugate) onto each reaction field of a clean reagent tray. Add all drops (reagent for a maximum of 50 slides) before continuing incubation. Use a stepper pipette. The labelled anti-human serum should be mixed with a pipette before use. To save time, conjugate can be pipetted onto separate reagent trays during incubation with the diluted serum. Incubate: Remove one BIOCHIP Slide from the PBS-Tween and within five seconds blot only the back and the long edges with a paper towel and immediately put the BIOCHIP Slide into the recesses of the reagent tray. Do not dry the areas between the reaction fields. Check for correct contact between the BIOCHIPs and liquids. Then continue with the next BIOCHIP Slide. From now on, protect the slides from direct sunlight. Incubate for 30 min at room temperature. Wash: Rinse the BIOCHIP Slides with a flush of PBS-Tween using a beaker and put them in a cuvette containing PBS-Tween for at least 5 min. 10 drops of Evans Blue (150 l) for each 150 ml phosphate buffer can be added for counterstaining. Embed: Place glycerol/PBS onto a cover glass drops of 10 l per reaction field. Use a polystyrene embedding template. Remove one BIOCHIP Slide from the PBS-Tween and dry the back and all four edges as well as the surface around, but not between, the reaction fields with a paper towel. Put the BIOCHIP Slide, with the BIOCHIPs facing downwards, onto the prepared cover glass. Check immediately that the cover glass is properly fitted into the recesses of the slide. Correct the position if necessary. Now proceed in the same way with the next BIOCHIP Slide. Evaluate: Read the fluorescence under the microscope.
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EUROIMMUN
slide BIOCHIPs reagent tray
Pipette: 10 l per field (3 x 3 mm) 25 l per field (5 x 5 mm) 70 l per field (7 x 9 mm)
;; ;; ;; ;;; ;; ;; ;; ;; ;;; ;; ;;; ;;;;

diluted samples
Incubate:
30 min
;; ;;;;; ;;
PBSTween
Wash:
1 s flush 5 min cuvette
Pipette: 10 l per field (3 x 3 mm) 20 l per field (5 x 5 mm) 60 l per field (7 x 9 mm)
;;; ;;; ;;; ;;; ;;; ;;; ;;; ;;; ;;; ;;; ;;; ;;; ;;; ;;; ;;;
labelled antibody ;;; ;;; ;;; ;;; ;;; ;;; ;;; ;;; ;;; ;;;
Incubate:
30 min
;; ;;;; ;;;; ;;;; ;;;; ;;

PBSTween
Wash:
Embed: 10 l per field (3 x 3 mm) 10 l per field (5 x 5 mm) 20 l per field (7 x 9 mm)
glycerol/PBS cover glass
Evaluate:
fluorescence microscopy
20x NEOFLUAR
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The Indirect Immunofluorescence Test, Performed Using the TITERPLANE Technique
EUROIMMUN
Recommended Serum Dilutions for Indirect Immunofluorescence Autoimmunity

Antibodies against acetylcholine receptor actin ADH-producing cells adrenal cortex alveolar basement membrane Substrate *skeletal muscle, monkey / heart, monkey Hepatitis Mosaic** nucl. supraopticus & paraventricularis, monkey adrenal gland, monkey lung, monkey / kidney, monkey IgA IgG IgM IgAGM 100 100 10 10
10
10 10 10 10
aquaporin-4 Neurology Mosaic asialoglycoprotein receptors *Hepatitis Mosaic** basic myelin protein (BMP) cerebellum, monkey / nerves, monkey / intestinal tissue, fetal monkey bile canaliculi Hepatitis Mosaic** bile duct epithelium Hepatitis Mosaic** BP180 BP230 brain: grey matter brain: white matter cANCA cartilage cell nuclei (ANA) CENP-F centromere cerebrum chondroitin sulfate collagen type VII collagenous connective tissue colon (epithelial cells) cornea cyclin I (PCNA) cyclin II (mitosin) cytoskeleton desmoglein 1+3 desmosomes dsDNA dsDNA elastin endocardium endomysium endoplasmatic reticulum endothelial cells endplates enterocytes eosinophilic granulocytes Dermatology Mosaic (EUROPLUS) Dermatology Mosaic (transfected cells) cerebellum, monkey / intestinal tissue, fetal monkey cerebellum, monkey / intestinal tissue, fetal monkey granulocytes (EOH), human / liver, monkey 10 trachea, fetal monkey HEp-2 cells / liver, monkey *HEp-2 cells / liver, monkey HEp-2 cells / liver, monkey gyrus precentralis, monkey trachea / cartilage, monkey oesophagus, monkey or tongue, monkey pancreas, monkey intestinal tissue, fetal monkey eye, monkey HEp-2 cells / liver, monkey HEp-2 cells / liver, monkey HEp-2 cells / liver, monkey transfected cells oesophagus, monkey or tongue, monkey 10
100
100 10 + 100 100 10 + 100
10 10 10 + 100 10 + 100 10 + 100 10 100 + 1000 + 10000 100 + 1000 + 10000 100 + 1000 + 10000 10 + 100 10 10 10 10 10 100 + 1000 + 10000 100 + 1000 + 10000 100 + 1000 + 10000 10 10 100 + 1000 + 10000 10 10
10 10 1 10 100 100 100 10
10 10 10 100 100 100
10
100
*HEp-2 cells / liver, monkey Crithidia luciliae stomach, rat / kidney, rat endocardium, monkey / intestinal tissue, fetal monkey liver, monkey 10 HEp-2 cells / liver, monkey lung, monkey *skeletal muscle, monkey / heart, monkey intestinal tissue, fetal monkey granulocytes (EOH) / liver, monkey
100 10 10 10
10 100 + 1000 + 10000 10 100 10 100 1 10 10 100 10 10 10 10 10 10
10 10
10 10 + 100 10 100 + 1000 + 10000 10 10
epidermal basement membrane oesophagus, monkey or tongue, monkey eye muscle eye, monkey fibrillarin (U3-nRNP) HEp-2 cells / liver, monkey filaggrin oesophagus, rat ganglion cells ganglion stellatum, monkey / intestinal tissue, fetal monkey gastric mucosa stomach, monkey gastrin-producing cells stomach (antrum), monkey / stomach (corpus), monkey glandula suprarenalis adrenal gland, monkey gliadin *intestinal tissue, fetal monkey / gliadin dots 10 glomerular basement membrane (GBM) kidney, monkey glutamate receptor type NMDA glutamic acid decarboxylase (GAD) goblet cells Golgi apparatus hair follicle transfected cells cerebellum, monkey / pancreas, monkey goblet cells (culture) HEp-2 cells / liver, monkey epidermis, monkey
10 10 10 10 10 + 100 10 100 + 1000 + 10000 10
10 100 10
10
*) In addition to the preferential analysis or as a plausibility check. **) Hepatitis Mosaic: liver, monkey / heart, monkey / HEp-2 cells / liver, rat / kidney, rat / stomach, rat.
12
EUROIMMUN
Antibodies against
Substrate
IgA
IgG
IgM
IgAGM
heart muscle skeletal muscle, monkey / heart, monkey 100 heart valves mitral valve, monkey 10 histones *HEp-2 cells / liver, monkey 100 + 1000 + 10000 Hu cerebellum, monkey / nerves, monkey / intestinal tissue, fetal monkey 10 + 100 hypothalamus nucl. supraopticus & paraventricularis, monkey 10 inner ear inner ear, rat or guinea pig 10 intercalated discs heart, monkey 100 intestinal epithelial tissue intestinal tissue, fetal monkey 10 10 intrinsic factor *stomach, monkey / intrinsic factor 10 10 Jo-1 *HEp-2 cells / liver, monkey 100 + 1000 + 10000 keratin oesophagus, monkey or tongue, monkey keratin, RA-associated (filaggrin) oesophagus, rat Ku *HEp-2 cells / liver, monkey labyrinth inner ear, rat or guinea pig lacrimal gland (excretory ducts and acini) lacrimal gland, monkey laminin lamins lipocytes liver membrane (LMA) liver-kidney microsomes (LKM) liver-pancreas antigen (LP) liver-specific protein (LSP) lymphocytes lysosomes M2 M3 M4 M5 M6 M7 M8 M9 medullated nerves melanocytes Mi-1 epidermis, monkey / lung, monkey / kidney, monkey HEp-2 cells / liver, monkey fat tissue, monkey Hepatitis Mosaic** Hepatitis Mosaic** Hepatitis Mosaic** / pancreas, monkey Hepatitis Mosaic** Lymphocytes HEp-2 cells / liver, monkey *Hepatitis Mosaic** *Hepatitis *Hepatitis *Hepatitis *Hepatitis *Hepatitis Mosaic** Mosaic** Mosaic** Mosaic** Mosaic** 10 10 100 + 1000 + 10000 10 10 10 100 + 1000 + 10000 10
100 10 100 10 + 100 10 10 100
100 10 10 100 10 10 10 100 10 100 100 100 100 10 + 100 100 100 100 100 100 100 100 100 100 10 + 100 10 100 100 100 10 10 + 100 10 + 100
10 + 100 100 + 1000 + 10000 100 + 1000 + 10000 100 + 1000 + 10000 100 + 1000 + 10000 100 + 1000 + 10000 100 + 1000 + 10000 100 + 1000 + 10000 100 + 1000 + 10000 100 + 1000 + 10000 10 100 + 1000 + 10000 100 + 1000 + 10000 100 + 1000 + 10000 10
*Hepatitis Mosaic** *Hepatitis Mosaic** cerebellum, monkey / nerves, monkey retina, monkey HEp-2 cells / liver, monkey
Mi-2 HEp-2 cells / liver, monkey mitochondria (AMA) kidney, rat / stomach, rat / M2 dots / HEp-2 cells mouth mucosa mouth mucosa myelin cerebellum, monkey / nerves, monkey myelin-associated glycoprotein (MAG) cerebellum, monkey / nerves, monkey myeloperoxidase (MPO) myocardium myolemma myosin native collagen nerves neuroendothelium neurofilaments NOR nRNP granulocytes (EOH), human / liver, monkey skeletal muscle, monkey / heart, monkey skeletal muscle, monkey / heart, monkey skeletal muscle, monkey / heart, monkey pancreas, monkey
10
10
10 + 100 100 10 + 100 100
1 100 10 + 100 100 10 10 + 100 10 + 100 10 + 100 100 100 10 1 10 10
cerebellum, monkey / nerves, monkey / intestinal tissue, fetal monkey 10 + 100 cerebellum, monkey / nerves, monkey / intestinal tissue, fetal monkey 10 + 100 cerebellum, monkey / nerves, monkey / intestinal tissue, fetal monkey 10 + 100 HEp-2 cells / liver, monkey 100 + 1000 + 10000 HEp-2 cells / liver, monkey 100 + 1000 + 10000 10 10 + 100 10 10 10
ovary ovary, monkey pANCA granulocytes (EOH), human / liver, monkey pancreas acini (Crohns disease autoantigen) pancreas, monkey pancreas islets pancreas, monkey (1st step: 18 hours) pancreas, excretory duct epithelium pancreas, monkey
10 10 10
*) In addition to the preferential analysis or as a plausibility check. **) Hepatitis Mosaic: liver, monkey / heart, monkey / HEp-2 cells / liver, rat / kidney, rat / stomach, rat.
13
EUROIMMUN

Antibodies against Substrate IgA IgG IgM IgAGM
parathyroid gland parathyroid gland, monkey 10 parietal cells stomach (corpus), monkey 10 10 parotid gland parotid gland, monkey 10 peripheral nerves cerebellum, monkey / nerves, monkey / intestinal tissue, fetal monkey 10 + 100 pituitary gland, anterior lobe pituitary gland, monkey 10 placenta placenta, human PM-1 HEp-2 cells / liver, monkey 100 + 1000 + 10000 proinsulin *pancreas, monkey 10 prostate prostate, monkey proteinase 3 (PR3) granulocytes (EOH), human / liver, monkey 10 10 + 100 Purkinje cell cytoplasm (Yo) RANA reticulin reticulocytes retina Ri ribosomal P-proteins ribosomes RNA polymerase RNA cerebellum, monkey Raji cells / HEp-2 cells intestinal tissue, fetal monkey bone marrow, monkey retina, monkey 10 + 100 10 10 10 10
10 10 + 100 10 10 + 100 10 10 100 10 10 1 10 + 100 10 10 10 10 10 + 100 100 100 100 100 10 10 + 100 100 100 100 100 100 10 100 10 + 100 100 100 100 100 10 + 100 10 10 10 10 10 10 10 10 10 100 10 10 100 1
10
cerebellum, monkey / nerves, monkey / intestinal tissue, fetal monkey 10 + 100 HEp-2 cells / liver, monkey 100 + 1000 + 10000 HEp-2 cells / liver, monkey 100 + 1000 + 10000 HEp-2 cells / liver, monkey 100 + 1000 + 10000 HEp-2 cells / liver, monkey 100 + 1000 + 10000 10 10 + 100 100 + 1000 + 10000 100 + 1000 + 10000 100 100 + 1000 + 10000 100 10 100 + 1000 + 10000
salivary glands (acini and excretory ducts) parotid gland, monkey sarcolemma skeletal muscle, monkey / heart, monkey Scl-70 HEp-2 cells / liver, monkey signal recognition particle (SRP) HEp-2 cells / liver, monkey skeletal muscle skeletal muscle, monkey / heart, monkey Sm smooth muscles (ASMA) spermatozoa spindle fibers spleen SS-A (Ro) SS-B (La) ssDNA striated muscles substantia nigra synovialis testis thrombocytes (bound antibodies) thrombocytes (free antibodies) thymus thyroglobulin thyroid colloid type II thyroid microsomes trachea tubular basement membrane U1-nRNP vasopressin-producing cells vestibular organ vimentin xANCA HEp-2 cells / liver, monkey stomach, rat / kidney, rat spermatozoa smear, human HEp-2 cells / liver, monkey spleen, monkey *HEp-2 cells / liver, monkey *HEp-2 cells / liver, monkey *HEp-2 cells / liver, monkey skeletal muscle, monkey / heart, monkey substantia nigra, monkey joint cartilage, monkey testis, monkey thrombocyte smear thrombocytes, human thymus, monkey thyroid gland, monkey or struma, human/TG thyroid gland, monkey or struma, human thyroid gland, monkey or struma, human trachea, monkey kidney, monkey *HEp-2 cells / liver, monkey nucl. supraopticus & paraventricularis, monkey inner ear, rat or guinea pig *HEp-2 cells / liver, monkey granulocytes (EOH, acetone, HCHO) / liver, monkey 10
10
100 + 1000 + 10000 100 + 1000 + 10000 100 + 1000 + 10000 100 10 10 10 10 10 10 + 100 10 10 + 100 10 10 100 + 1000 + 10000 10 10 100 + 1000 + 10000 10 + 100
10
*) In addition to the preferential analysis or as a plausibility check.

14
EUROIMMUN
Antibodies against Adenovirus (type 3) Afipia felis Bartonella henselae Bartonella quintana Bordetella parapertussis Bordetella pertussis Borrelia afzelii Borrelia burgdorferi sensu stricto (strains CH and USA) Borrelia garinii Borrelia OspC Borrelia VlsE Campylobacter coli Campylobacter jejuni Candida albicans Candida glabrata Candida krusei Candida parapsilosis Candida tropicalis Chikungunya virus Chlamydia pneumoniae (IFA) Chlamydia Chlamydia Chlamydia Chlamydia CMV pneumoniae (MIF) trachomatis (IFA) trachomatis (MIF) psittaci (MIF)
IgA 10 + 100 100
IgG
IgM
10 + 100 10 + 100
10 + 100 10 100 + 1000 10 100 + 320 + 1000 100 100 + 320 + 1000 100 100 + 1000 100 + 1000 100 + 1000 100 + 1000 100 + 320 + 1000 10 100 + 320 + 1000 10 100 + 320 + 1000 10 10 100 100 + 1000 1000 1000 + 10000 1000 + 10000 1000 + 10000 1000 + 10000 1000 + 10000 10 + 100 100 + 1000 100 100 + 320 + 1000 100 100 100 + 1000 100 + 1000 100 + 1000 10 + 100 + 1000 10 + 100 100 + 320 + 1000 100 + 1000 100 + 1000 1000 + 10000 100 + 1000 10 + 100 10 + 100 100 + 1000 + 10000 10 + 100 10 + 100 100 100 + 320 + 1000 320 100 + 1000 10 + 100 10 + 100 10 + 100 10 + 100 10 + 100 + 1000 32 + 100
100 320 100 + 1000 100 + 1000 100 + 1000 100 + 1000 100 + 1000 100 10 100 10 10 100 10 + 100 10 10 + 100 100 10 + 100 100 + 1000 100 100
10 100 100 100 100 100 100 10 + 100 10 10 10 10 10 100 10 10 + 100 10 + 100 100 10 100 + 1000 10 10
Coxsackie virus types A7, A9, A16, A24, B1 to B6 Dengue virus EBV-CA, -EA EBNA Echinococcus granulosus Echo virus (type 7, 19) Hanta virus Haemophilus influenzae Helicobacter pylori HIV-1/2 HHV-6 HSV-1/2 Influenza virus type A Influenza virus type B Klebsiella pneumoniae Legionella pneumophila (all serotypes) Leishmania Listeria monocytogenes (type 1/2a, 4b) measles virus mumps virus Mycoplasma hominis Mycoplasma pneumoniae Parainfluenza virus types 1 - 4 Plasmodium falciparum/vivax rubella virus RSV Saccharomyces cerevisiae TBE virus Toxoplasma gondii Treponema pallidum Ureaplasma urealyticum VZV West-Nile virus Yersinia enterocolitica O:3; O:4; O:6; O:9
10 10 10 100
10 10 10 10 100
100 100 10 10 10 10 10 + 100
100 100 10 10 10 10 10
10 + 100 10 + 100 10 + 100 10 10 + 100 + 1000 10 100 1000 100 10 10 + 100 + 1000 10 + 100 16+64+256 16+64+256+1028... 16+64+256 10 10 10 10 + 100 10 + 100 10 + 100 10 + 100 10 + 100 + 1000 100 10 10 10 10 + 100 10 + 100
15
Recommended Serum Dilutions for Indirect Immunofluorescence Infectious Serology
EUROIMMUN
Diagnostically Relevant Autoantibodies

Systemic Autoantibodies against
Ig- AGM A G M 151 152 1572 1572 1590
1590
1590
162 171 120 121 1010 1030
BASIS SPECTRUM ANA (cell nuclei) IF global testing ANA profile differentiation dsDNA-NcX ELISA dsDNA IFT SLE specific ENA ProfilePlus ELISA 1 nRNP/Sm, Sm, SS-A, SS-B, Scl-70, Jo-1 ENA ProfilePlus ELISA 2 rib. P proteins, RNP/Sm, Sm, SS-A, SS-B, Scl-70, Jo-1, CENP B SLE Profile ELISA (dsDNA, histones, rib. P prot., nRNP/Sm, Sm, SS-A, SS-B, Scl-70) AMA (mitochondria) ASMA (smooth muscle) cANCA* (granulocytes) Wegeners dis. pANCA* (granulocytes) vasculitis autoantibody profile 10 IF substrates autoantibody profile 30 IF substrates
Ig- AGM A G M ANA DIAGNOSTICS, WESTERNBLOT 1520 PM-Scl, CENP A/B, Ku 86 and 72 kDa, M2 74 kDa, RNP 70 kDa, RNP A/C, Sm B/B/D, SS-A 60 and 52 kDa, SS-B 52, 47, 44 and 43 kDa, ribosomal P proteins P0/P1/P2, Scl-70, Jo-1)
Ig- AGM A G M SYSTEMIC LUPUS ERYTHEMATOSUS (SLE) 151 ANA (cell nuclei) IF global testing 1574 nucleosomes SLE specific 1572 dsDNA ELISA 1572 dsDNA IFT (C. luciliae) SLE specific 1572 dsDNA RIA 159 ENA PoolPlus ELISA 1591 U1-nRNP 1593 Sm SLE specific 1595 SS-A (Ro) 159s SS-A 52 kDa: recombinant 159t SS-A 60 kDa: recombinant 1597 SS-B (La) 1640 ribosomal P proteins SLE specific 1605 Ku 1601 cyclin I (PCNA) 156 histones (global testing) 1576 ssDNA (single-stranded DNA) 121 pANCA* (granulocytes) vasculitis
Ig- AGM A G M 151 1595 159s 159t 1597 SJGRENS SYNDROME ANA (cell nuclei) IF global testing SS-A (Ro) SS-A 52 kDa: recombinant SS-A 60 kDa: recombinant SS-B (La)
Ig- AGM A G M POLYMYOSITIS, DERMATOMYOSITIS 151 ANA (cell nuclei) IF global testing 1530 Myositis Profile 3 EUROLINE (Mi-2, Ku, PM-Scl100, PM-Scl75, SRP, Jo-1, PL-7, PL-12, OJ, EJ, Ro-52) 1661 Jo-1 1662 PL-7 1663 PL-12 1664 OJ 1665 EJ 1666 SC 1667 KS 1584 PM-Scl (PM-1) 1616 SRP (signal recognition particle) 159s SS-A 52 kDa: recombinant 1605 Ku 1607 Mi-2 1635 serotonin ab 1636 PMR (polymyalgia rheumatica factor)
Ig- AGM A G M 1505 151a 1814 1508 1219 151 1604 121 148 194 1947
Ig- AGM A G M 2011 2012 2013 2014 2031 2034 213 2171 2191
Ig- AGM A G M 1811 1813 1814 1815
Ig- AGM A G M 1612 1613 1614 1615 1617 1641 1642 1643 165 1651 1652 1653 1654 1655 1656 1659 194 1947 1950 196
Ig- AGM A G M 1821 1822 1824 1572 1818
OTHER AUTOANTIBODIES centrioles MSA-1 (NuMa, spindle fibres) MSA-2 (midbody) MSA-3 (chromosome-ass. antigen) centromer F protein (CENP-F) ribosomes Golgi apparatus lysosomes cytoskeleton actin vimentin cytokeratin tropomyosin vinculin desmin laminin (basal membranes) collagen (global testing) collagen type VII elastin vessel endothelium
THERAPY CONTROL interferon alpha*** interferon beta erythropoetin dsDNA RIA CIC-C1q ELISA
Ig- AGM A G M ANTI-PHOSPHOLIPID SYNDROME (APS) 1621 cardiolipin 1632 -2-glycoprotein 1 1631 lupus anticoagulant (plasma)** 162a phosphatidylserine
(RHEUMATOID) ARTHRITIS CCP (cyclic citrullinated peptides) Sa RF IgM (class. rheumatoid factor) filaggrin (RA keratin) GS ANA (granulocyte specific ANA) ANA (cell nuclei) IF global testing RANA (rheum. arthritis nuclear antigen) pANCA* (granuloc.) RF-ass. vasculitis cartilaginous subst. polychondritis collagen (global testing) collagen type VII
FURTHER RHEUMATOID RELEVANT ANALYSES anti-streptolysin anti-streptokinase anti-streptodornase anti-DPNase (anti-NADase) anti-staphylolysin anti-hyaluronidase Borrelia burgdorferi Yersinia enterocolitica O:3 Chlamydia trachomatis
IMMUNOGLOBULINS: ANTIhuman IgA human IgE human IgG human IgM
Ig- AGM A G M 151 1599 1584 1611 1611 1582 1583 1585 1586 1587
PROGRESSIVE SYSTEMIC SCLEROSIS ANA (cell nuclei) IF global testing Scl-70 (DNA topoisomerase I) PM-Scl (PM-1) centromeres centromere B protein (recombinant) U3-nRNP (fibrillarin) RNA polymerase I, II, III 7-2-RNP (To) 4-6-S-RNA NOR (nucleolus organizer region)
Ig- AGM A G M SHARPS SYNDROME MCTD 1591 U1-nRNP 151 ANA (cell nuclei) IF global testing
Ig- AGM A G M CREST SYNDROME 1611 centromeres 1611 centromere B protein (recombinant)
1818
CIRC. IMMUNE COMPLEXES C1q ELISA
Grey boxes: standard analysis
*) ANCA diagnostics in acute cases within one hour, at any hour
**) Use special procedure for taking sample(s)
***) Send frozen sample(s)
16
EUROIMMUN
Diagnostically Relevant Autoantibodies

Ig- AGM A G M AUTOANTIBODY PROFILE 30 IF substrates (BIOCHIPs) 1030 Ig- AGM A G M 1015 1012 1013 1014 1011 1016 1017 Ig- AGM A G M 1021 1022 1023 1024 1025 1026 147 Ig- AGM A G M 1051 1053 1061 1062 1081 105 104 1021 1012 1361 1361 1091 1092 1011 1052 107 110 Ig- AGM A G M 1621 1060 1081 1086 1091 107 1401 Ig- AGM A G M 1501 1495 1496 1502 1502 1502 135 1503 1509 191 3011 1502 1504 150h Ig- AGM A G M 1178 1177 1171 1172 1173 1174 1175 1176 1119 120 151 Ig- AGM A G M 124 1209 1221 1231 1232 1361 1361 THYROID GLAND TRAb (TSH receptors) TPO ab (thyroidea peroxidase) TAb (thyroglobulin) colloid antigen II ab MAb (microsomes) T3 ab T4 ab DIABETES MELLITUS ICA (islet cell antibodies) GAD (glutamic acid decarboxylase) IA-2 (tyrosine phosphatase) insulin ab human insulin receptor glucagon-producing cells lipocytes (POLY-)ENDOCRINOPATHY adrenal cortex Addisons dis. 21-hydroxylase Addisons dis. ovary: theca cells ovary: corpus luteum testis: Leydig cells steroid hormon-producing cells parathyroid gland ICA (islet cell antibodies) TPO ab (thyroidea peroxidase) H+/K+-ATPase ab ELISA PCA (parietal cells) pituitary gland: anterior lobe pituitary gland: posterior lobe MAb (thyroid microsomes) adrenal medulla placenta VPZ (vasopr.-prod. cells) D. insipudus INFERTILITY cardiolipin ovary: theca c., c. luteum, z. pellucida testis: Leydig cells spermatozoa pituitary gland: anterior lobe placenta prostate EPIDERMIS desmosomes pemphigus desmoglein 1 pemphigus desmoglein 3 pemphigus epiderm. basal membr. pemphigoid BP180 bullous pemphigoid BP230 bullous pemphigoid oral mucosa Behets/ Crohns dis. basal membrane (urinary bladder) epidermal keratin endomysium GSE, Duhrings dis. gliadin GSE, Duhrings dis. herpes gestationis factor melanocytes hair follicle EYE recoverin tunica choroidea chron. chorioretinitis cornea retina lens oculi corpus ciliare eye muscles retro bulbar connective tissue other: CV2 (oligodendrocytes) cANCA* (granulocytes) Wegeners dis. ANA (cell nuclei) IF global testing IMMUNOHAEMATOLOGY erythrocytes (global testing) granulocyte membrane lymphocytes thrombocytes: indirect test (free ab) thrombocytes: direct test (bound ab)** H+/K+-ATPase ab ELISA PCA (parietal cells) Ig- AGM A G M 120 1201 1202 121 1211 1212 1213 1215 120 151 195 196 Ig- AGM A G M 120 121 125 1251 151 1572 1252 1271 WEGENERS DISEASE, VASKULITIS* cANCA IFT* granuloc. Wegeners dis. PR3 (proteinase 3) BPI (CAP 57) pANCA IFT* granulocytes vasculitis MPO (myeloperoxidase) elastase cathepsin G lactoferrin ANCA Profile ELISA PR3, MPO, elastase, cath. G, BPI, lactoferrin ANA (cell nuclei) IF global testing elastin vessel endothelium Ig- AGM A G M LIVER, BILIARY DUCTS 130 Liver Ab IFT global testing, 6 BIOCHIPs Autoimmune Liver Dis. Ab Profile 130 EUROLINE AMA-M2, 3E (BPO), Sp100, PML, gp210, LC-1, LKM-1, SLA/LP, Ro-52 1302 1651 151 1307 132 1321 1322 1323 1303 171 1301 1304 Autoimmune hepatitis (AIH) SLA/LP (soluble liver antigen) F-actin ANA (cell nuclei) IF global testing LC-1 (liver cytosol) LKM (liver kidney microsomes) LKM-1 ELISA LKM-2 LKM-3 ASGPR (asialoglycoprotein receptors) ASMA (smooth muscles) LSP (liver-specific protein) LMA (liver cell membrane) Primary biliary cirrhosis (PBC) AMA (mitochondria) AMA-M2 (PDH + BPO) AMA-M4 (sulfitoxidase) AMA-M9 (glycogen phosphorylase) Sp100 (nuclear dots) GP210 (nuclear membrane, lamin) Primary-sclerosing cholangitis (PSC) pANCA (granulocytes) further antibodies bile ducts bile canaliculi coilin; P80 (few nuclear dots) STOMACH, INTESTINE PCA (parietal cells) atroph. gastritis H+/K+-ATPase ab atroph. gastritis intrinsic factor ab vit. B12 deficiency gastrin (G) cells pancreas acinus cells Crohns dis. CUZD1 Crohns dis. GP2 Crohns dis. Saccharomyces cerev. Crohns dis. pankreas secretion Crohns dis. mouth mucosa Behets/ Crohns dis. intestinal goblet cells ulc. colitis pANCA (granulocytes) ulc. colitis enterocytes Crohns dis. , ulc. colitis endomysium GSE, Duhrings dis. transglutaminase GSE, Duhrings dis. deamidated gliadin (Z-AGFA) GSE reticulin GSE, Duhrings dis. EXOCRINE GLANDS, PANCREATITIS, SJGRENS SYNDROME exocrine pancreas pancreas acini pancreas excretory ducts salivary glands (parotid gland) parotid gland acini parotid gland excretory ducts lacrimal gland Sjgrens syndrome ANA (cell nuclei) IF global testing SS-A (Ro) SS-B (La) ssDNA (single-stranded DNA) further antibodies prostate mamma HEART AMA-M7 (myocard-specific) heart muscle heart: intercalated disk heart: myolemma
Organ-/Tissue-Specic Autoimmunity: Autoantibodies against
KIDNEY, LUNG cANCA IFT* granuloc. Wegeners dis. pANCA IFT* granulocytes vasculitis kidney IF global testing GBM ELISA glomerular basal membrane ANA (cell nuclei) IF global testing dsDNA IFT 162 TBM (tubular basal membrane) 1622 lung alveolar basal membrane 1624 1629 Ig- AGM A G M NERVOUS SYSTEM 1603 111 Neuronal Ab IFT global testing 1608 1111 Paraneoplastic neurol. syndromes Neuronal Antigens Profile 2 EUROLINE amphiphysin, CV2.1***, PNMA2 (Ma-2), Ri, Yo, Hu Tr (Purkinje cell cytoplasm) Yo (Purkinje cell cytoplasm; PCA-1) PCA-2 (Purkinje cell cytoplasm) Ri (neurone nuclei; ANNA-2) Hu (neurone nuclei; ANNA-1) NMDA receptors Ma1/Ma2 (neurone nuclei; Ta) CV2 (oligodendrocytes) GAD stiff-person syndr. amphiphysin stiff-person syndr. AGNA (anti-glia nuclear antigen; SOX-1) ANNA-3 CRMP-5 (oligodendrocytes) further parameters aquaporin-4 neuromyelitis optica NMO-IgG neuromyelitis optica MOG (myelin-oligodendroc. glykoprot.) substantia nigra myelin MBP (myelin-basic protein) MAG (myelin-assoc. glycoprotein) myelin of peripheral nerves neuroendothelium neurofilaments GFAP (glial fibrillary acidic protein) non-medullated nerves astrocytes basal ganglia ganglion stellatum plexus myentericus synaptophysin ganglioside profile GM1, GM2, GM3, GD1a, GD1b, GT1b, GQ1b GM1 GM2 GM3 GD1a GD1b GT1b GQ1b neurofascin ANA (cell nuclei) IF global testing Borrelia burgd.: serum CSF SKELETAL MUSCLE, THYMUS acetylcholine receptors M. gravis MuSK M. gravis calcium channels type N LEMS calcium channels type PQ LEMS potassium channel ab (VGKC) thymus M. gravis, thymoma titin M. gravis skeletal muscle M. gravis sarcolemma myosin 121
1112 1113 1114 1115 1116 112d 1117 1119 1022 112e 112a 112b 112c 1154 1153 1156 1111 1121 1122 1123 1124 1126 1127 1128 1129 112f 112g 112h 112i 112j 1130 1131 1132 1133 1134 1135 1136 1137 1155 151 213 Ig- AGM A G M 1435 1434 1437 1438 1439 144 1431 143 1432 1436
1305 1306 1609 Ig- AGM A G M 1361 1361 1362 1366 1391 1391 1392 2841 1392 135 1381 121 1382 191 191 3011 192 Ig- AGM A G M 139 1391 1393 142 1421 1423 141 151 1595 1597 1576 1401 1406 Ig- AGM A G M 1627 146 1462 1463
Ig- AGM A G M LIPODYSTROPHY lipocytes 147 Ig- AGM A G M ANTIBODIES AGAINST ANIMAL IgG HAMA (human anti-mouse IgG) 17 3811
Grey: standard *) ANCA diagn. in acute cases within 1 h **) Special procedure for taking sample ***) CV2 partial protein, which only contains the N-terminally localised epitopes of the antigen
EUROIMMUN
Antibodies for Infectious Serology

Antibodies against
Respiratory tract Exanthema Lymphadenitis CNS Myocarditis Infectious arthritis Gastrointestinal tract Associated hepatitis Ophthalmology Otitis STD Pregnancy IFT ELISA Westernblot EUROLINE Respiratory tract Exanthema Lymphadenitis CNS Myocarditis Infectious arthritis Gastrointestinal tract Associated hepatitis Ophthalmology Otitis STD Pregnancy IFT ELISA Westernblot EUROLINE
Ig- AGM A G M 219a 219b 219d 2055 2050 2131 2132 2133 2134 2092 2091 2192 2191 2040 2070 2080 2101 216a 2167 2166 2165 2168 2169 2150 2140 2201 2202 2060 2111 2205 2170 Ig- AGM A G M 2320 2231 2261 2264 2410 Grey: standard analyses
SI I UK A /
BACTERIA A-Z Apia felis* Bartonella henselae Bartonella quintana Bordetella parapertussis Bordetella pertussis Borrelia afzelii Borrelia burgd. s. stricto CSF diagnostics Borrelia burgd. (USA) Borrelia garinii Campylobacter coli Campylobacter jejuni Chlamydia pneumoniae Chlamydia trachomatis Diphtheria toxoid Haemophilus inuenzae Helicobacter pylori Klebsiella pneumoniae Legionella bozemanii Legionella dumofi Legionella gormanii Legionella jordanis Legionella longbeachae Legionella micdadei Legionella pneumophila Serotypes .................. Listeria monocytogenes 1/2a 4b Mycoplasma hominis Mycoplasma pneumoniae Tetanus toxoid Treponema pallidum CSF diagnostics Ureaplasma urealyticum Yersinia enterocolitica O:3 O:4 O:6 O:9 PARASITES A-Z Echinococcus gran. Leishmania donovani Plasmodium vivax Plasmodium falciparum Toxoplasma gondii Avidity determination *) Currently not available as IVD in the European Union
Ig- AGM A G M 2680 2730 2570 275a 2791 2795 2793 2531 2532 2511 2512 2536 2691 2692 2610 2630 2720 2670 2590 2661 2650 Ig- AGM A G M 2861 2862 2863 2865 2864 2841
VIRUSES A-Z Adenovirus type 3 Coxsackievirus type B1 B2 B3 B4 B5 B6 A7 A9 A16 A24 Cytomegalovirus Avidity determination Echovirus type 7 Epstein-Barr virus capsid Ag (EBV-CA) Avidity determination Epstein-Barr virus early Ag (EBV-EA) Epstein-Barr virus nuclear Ag (EBNA) HSV-1 HSV-2 HIV-1* HIV-2* Human herpes virus 6 (HHV-6) Inuenza virus type A H1N1 H3N2 Inuenza virus type B Measles virus CSF diagnostics Mumps virus CSF diagnostics Parainuenza virus type 1 2 3 4 Respiratory syncytial virus (RSV) Rubella virus Avidity determination CSF diagnostics TBE virus Varicella zoster virus Avidity determination FUNGI A-Z Candida albicans Candida glabrata Candida krusei Candida parapsilosis Candida tropicalis Saccharom. cerevisiae
18
EUROIMMUN
Antibodies for Allergology

GLOBAL TEST 3840 Determination of total IgE (ELISA) 3410 3410 INHALATION 3110 3110 3111 3112 3113 Allergy Profile Inhalation (g1, g3, g6, g12, t2, t3, t4, t7, w1, w6, w9, d1, d2, e1, e2, e3, m1, m2, m3, m6, CCD) Allergy Profile Inhalation 2 (g6, g12, t2, t3, t4, w6, w9, d1, d2, e1, e2, e3, e6, e82, e84, es4, m1, m2, m3, m6, CCD) Allergy Profile Pediatric Inhalation (g6, g12, t2, t3, t4, w6, w8, w9, d1, d2, e1, e2, e3, e6, e82, e84, m1, m2, m3, m6, CCD) Allergy Profile Mediterranean Inhalation (g2, g6, t3, t4, t9, t11, t23, t210, w1, w6, w9, w19, d1, d2, d70, e1, e2, e3, m2, m6, CCD) Allergy Profile Inhalation "South East Asia" (ts19, t104, t19, t223, gs1, ds1, i6, u134, e1, e2, es172, e6, e71, e82, e84, ms1, ms4, m5, m12, m45, CCD) Allergy Profile Inhalation "China" (gs23, ts21, t3, t8, t11, t12, t14, t70, ws18, w1, w6, w9, es1, d1, d2, i6, ms5, m1, u73, u80, CCD) Allergy Profile Inhalation "Middle East" (g1, g6,g12, t2, t3, t7, t9, w1, w6, w8, d1, d2, i6, e1, e84, m1, m2, m3, m5, m6, CCD) Allergy Profile Inhalation "Gulf" (g6, g12, t2, t3, t7, t9, w1, w6, d1, d2, i6, e1, e2, e3, e17, m1, m2, m3, m5, m6, CCD) All. Profile Inhalation Mix-Screen Turkey 1 (ts23, ts24, ts25, gs12, gs15, gs21, ws17, ws18, ws19, ws20, ms11, ms12, CCD) Allergy Profile Inhalation Turkey 1 (gs12, gs15, gs21, g12, ts23, ts24, t9, t70, ws18, ws19, ws20, d1, d2, i6, es2, es172, e1, e2, e3, e4, e80, e81, e84, ms11, ms12, m1, m2, m3, m6, CCD) Allergy Profile Inhalation Turkey 2 (m1, m2, m3, m6, ds1, i6, e1, e2, e3, e4, e81, e84, CCD) Allergy Profile Inhalation "India" (g6, g12, g20, t18, w4, w27, w29, ds1, d2, i6, e1, e2, e11, e85, m3, m37, u81, u126, u129, u140, CCD) 3411 3411 3414 3415 3416 3417 3418 3419 3420 3420 FOOD Allergy Profile Food (f1, f75, f2, f45, f4, f5, f9, f13, f14, f17, f20, f49, f84, f237, f25, f31, f35, f85, f3, f23, CCD) Allergy Profile Food 2 (f1, f75, f2, f78, f4, f5, f14, f10, f13, f17, f20, f49, f84, f95, f25, f31, f35, f85, f3, f23, CCD) Allergy Profile Food "South East Asia 1" (f1, f75, f2, f4, f9, f10, f14, f13, f17, f63, f64, f83, fs10, fs14, f23, f24, f80, f179, f105, f336, CCD) Allergy Profile Food "South East Asia 2" (f1, f75, f2, f4, f9, f10, f14, f13, f17, f63, f340, f83, fs10, fs14, f23, f24, f80, f179, f105, f336, CCD) Allergy Profile Food "China" (f1, f2, f4, f7, f27, f88, fs35, f13, f14, fs40, f25, f292, fs42, f23, f234, f3, f41, f56, fs41, fs77, CCD) Allergy Profile Food "Middle East" (f1, f75, f2, f78, e204, f4, f14, f45, f13, f17, f20, f33, f49, f92, f25, f31, f85, f48, f88, f89, CCD) Allergy Profile Food "Gulf" (f1, f75, f2, f105, f4, f14, f45, fs36, f13, f29, f33, f44, f93, f25, f31, f48, f83, f88, f3, f23, CCD) Allergy Profile Food "Cyprus 1" (f1, f2, f75, f76, f77, f78, f81, f4, f45, f5, f14, f7, f79, f9, f10, f13, f144, f17, f20, f49, CCD) Allergy Profile Food "Cyprus 2" (f177, f23, f234, f24, f258, f3, f37, f40, f41, f80, f48, f108, f132, f212, f25, f292, f31, f35, f47, f96, CCD) Allergy Profile Food "Cyprus 3" (f26, f63, f83, f88, f237, f29, f32, f328, f33, f44, f49, f72, f84, f95, f281, f86, f89, f90, f105, f93, CCD) Allergy Profile Food Mix Screen Turkey 1 (f245, fs8, fs50, fs38, fs37, fs46, fs47, fs48, fs49, fs45, fs10, fs16, CCD) Allergy Profile Food Turkey 1 (f1, f75, f2, f169, f78, f4, f79, f9, f14, f10, f13, f17, f144, u87, f222, f73, f33, f44, f49, f92, f84, f146, f328, f25, f31, f35, f48, f95, f97, f122, f132, fs14, fs10, fs43, f83, CCD) Allergy Profile Food Turkey 2 (f3, f21, f206, f437, f438, f436, f71, f177, f324, f27, f83, f88, CCD) Allergy Profile Food Turkey 3 (f1, fs51, f14, fx20, f13, fs35, f49, f44, f25, f31, fs10, fs16, CCD) Allergy Profile Food "India" (f2, f75, f168, f4, f9, f14, f13, f36, f49, f50, f35, f38, f48, f244, f83, f89, f74, f240, f23, f24, CCD) 3710 3710 3711 ATOPY, POLLEN-ASSOCIATED FOOD ALLERGIES Allergy Profile Atopy (g6, g12, t3, w6, d1, e1, e2, e3, m2, m6, f1, f2, f3, f4, f9, f14, f17, f31, f35, f49, CCD) Allergy Profile Atopy 3 (g6, t3, t4, w6, d1, d2, e1, e2, e3, m2, m3, f1, f75, f2, f3, f4, f13, f14, f31, f49, CCD) Allergy Profile Pollen-Food Cross Reactions (g6, t3, w6, f4, f5, f13, f17, f20, f48, f89, f271, f275, f44, f49, f348, f237, f328, f31, f35, f85, CCD) Allergy Profile Pediatrics (gx, t3, w6, d1, e1, e2, e3, m2, m6, f1, f75, f3, f2, f76, f77, f78, e204, f4, f9, f14, f13, f17, f31, f35, f49, CCD) Allergy Profile Atopy "China" (ts20, w1, w6, ds1, h1, e1, e2, i6, ms1, u80, f1, f2, f13, f14, f27, f88, fs33, fs34, f23, f234, CCD) Allergy Profile Atopy "China 3" (ts22, w6, us1, ds1, es1, h1, ms5, f245, fs9, fs43, fs56, fs34, fs44, CCD) Allergy Profile Atopy Mix Screen Turkey 1 (hs12, ms1, gs2, ws21, ts26, ts25, fx5, fx20, es5, es6, es172, es2, CCD) Allergy Profile Atopy Turkey 1 (d1, m2, es5, g6, t3, w6, f1, f2, f13, f14, f4, fs16, CCD)
Allergen Profiles: Antibodies of class IgE against
3712
3713 3713 3719 3719
3116 3117 3118 3119 3119
3119 3120
3420 3420 3421
INSECT VENOMS 3720 Allergy Profile Insect Venoms (i1, i3, CCD)
Allercoat 6 System: Antibodies of class IgE against 600 different allergens of the areas animal allergens, environmental allergens, food, grasses, herbal and flower pollen, house dust, insects, mites, moulds, parasites, pharmaceutical drugs, trees.
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EUROIMMUN
EUROIMMUN Microplate ELISA

stain

substrate POD POD-labelled anti-human antibody POD chromogen
Polystyrene microplate strips coated with purified, biochemically characterized antigens are used as solid phase containing bound antigens. If the sample is positive, specific antibodies in the diluted serum sample attach to the antigens coupled to the solid phase. In a second step, the attached antibodies are detected with peroxidase-labelled anti-human antibodies. In a third step, the bound antibodies are made visible using a chromogen/ substrate solution which is capable of promoting a color reaction. The intensity of the color produced is proportional to the concentration of antibodies in the serum sample. Monospecific ELISA (enzyme immunoassays with a single antigen) provide a quantitative in vitro assay for the detection of antibodies. Profile ELISA provide a semiquantitative in vitro assay for the detection of different antibodies on a single microplate strip. The solid phase of Pool ELISA is coated with an antigen mixture for the semiquantitative detection of antibodies whose specificity must be subsequently investigated by monospecific assays.

antigen-coated microplate well
Reliable and Economical Calibration/Evaluation

In the case of a quantitative ELISA, calibration is performed using three calibration sera. Calibration serum 1: upper limit of the measurement range Calibration serum 2: upper limit of the normal range (cut-off value) Calibration serum 3: negative Only three wells are required for calibration, followed by serum samples. There is no need to incubate blank values or duplicate determinations. Semiquantitative ELISA are performed using only one calibrator. The calibration is performed in relative units per milliliter (RU/ml) or, if an international reference serum exists, in international units per milliliter (IU/ml). Each test can be optionally performed using a positive or negative control serum included in the test kit. Kit-specific reference ranges are provided for each calibrator and control serum. All calibrations can be easily performed with the usual ELISA software.
Easy, Quick and Economical Handling

A B C D E F G H
1 2 3 4 5 6 7 8 9 10 11 12
Microplate strips containing break-off wells (except Profile ELISA), each marked with an antigen abbreviation to avoid confusion of wells. Reagents ready for use (wash buffer: concentrate). Reagents are color-coded to ensure positive identification. dark red: calibration serum 1 orange: anti-human IgA POD-conj. red: light red: dark blue: calibration serum 2 calibration serum 3 pos. control serum green: red: turquoise: anti-human IgG POD-conj. anti-human IgM POD-conj. anti-human IgGM POD-conj.
YL. H. P
green: neg. control serum yellow: anti-human IgAGM POD-conj. light blue: sample buffer The sample buffer for infectious serology ELISA (detection of antibodies of class IgM) already contains an IgG/RF absorbent. Several tests can be combined on one and the same microplate, since the incubation times (30 min; 30 min; 15 min) are identical for all ELISA. Incubation at room temperature. Compatible with all commercial washer and reader systems.
H. PYL.
H. PYL.
H. PYL.
H. PYL.
H. PYL.
YL. H. P
YL. H. P
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EUROIMMUN
Determination of Low-Avidity Antibodies

An alternative principle for the serological diagnosis of fresh infections has been established by investigating the antibody avidity. The first reaction of the immune system following an infection is the formation of low-avidity antibodies. As the infection proceeds, increasingly antigen-adapted IgG is formed, and avidity grows. As long as high-avidity IgG is not yet detected in the serum, it can be assumed that the infection is still in an early stage. To identify low-avidity antibodies in a patients serum, two microplate ELISA are performed in parallel: one test is carried out in the conventional way, the other one includes urea treatment between incubations with patients serum and peroxidase-labelled anti-human IgG, resulting in the detachment of low-avidity antibodies from the antigens. Low-avidity antibodies are present if the ELISA extinction is significantly reduced by urea treatment. For an objective interpretation, the relative avidity index (RAI) can be calculated out of the measured values with and without urea incubation. Serum dilution 1 : 100, conjugate class anti-human IgG, POD-labelled. 3-point calibration, quantitative (IgG). The following test kits for avidity determination are available: Toxoplasma gondii, CMV, rubella virus, VZV, West-Nile virus, EBV-CA.
1 2 3 4 5 6
10
Number of Patients
RAI =
Ewith urea Ewithout urea Primary Infection Previous Infection
25 50 75 100 Relative Avidity Index (RAI) in %
Avidity of antibodies against EBV-CA (IgG)
Antibody Determination in CSF

Indication: local infections of the brain. CSF dilution 1 : 2, serum dilution 1 : 404. Conjugate classes anti-human IgG or IgM, POD-labelled. Easy to conduct: ready-for-use reagents. 4-point calibration, quantitative. Identical incubation conditions and times (room temperature; 60 min / 60 min / 15 min): all EUROIMMUN ELISA for CSF diagnostics can be combined without difficulty on one and the same microplate. The antibody concentration in the patients serum is determined in parallel to the antibody concentration in CSF on one and the same microplate. The CSF/ serum quotient CSQpath.-spec. is calculated from both measured values. An intrathecal synthesis of specific antibodies is present if the CSF/serum quotient of the specific antibodies CSQpath.-spec. is significantly higher than the CSF/serum quotient of the whole IgG (CSQtotal) or if necessary the CSQlim.. The relation of both values indicates the relative CSF/serum quotient CSQrel. (synonym: antibody specificity index, ASI). Interpretation of results (according to the recommendations of Prof. Reiber): standard range CSQrel. < 1,3: CSQrel. 1,3 1,5: borderline range CSQrel. > 1,5: Indication of pathogen-specific antibody production in the CNS For the automatic calculation of the CSQrel. EUROIMMUN provides a specific Excel table free of charge. Highest sensitivity, specificity and reproducibility. Antibody concentrations in the serum and CSF are determined within the linear range of the test. The following test kits for CSF diagnostics are available: Borrelia burgdorferi, Toxoplasma gondii, HSV-1, HSV-2, HSV-1/2 Pool, CMV, rubella virus, measles virus, mumps virus, VZV, TBE, EBV-CA. All test systems for CSF diagnostics can also be used only for serology. Perfectly adapted for the automated incubation in incubation devices.
A B C D E F G H
CA-D
CA CB CC CD C1 1:2 S1 1:401 C2 1:2 S2 1:401
C3 1:2 S3 1:401 C4 1:2 S4 1:401 C5 1:2 S5 1:401 C6 1:2 S6 1:401

C
CSF calibrators
CSF
serum
ELISA incubation scheme
Table-based evaluation of the CSQrel.
Reference range for normal values, intact blood-CSF barrier function

CSQtotal
Blood-CSF barrier dysfunction, no Ig production in CNS Blood-CSF barrier dysfunction, additional Ig production in CNS Clear Ig production in CNS, no disturbance in blood-CSF barrier function Error in blood extraction or analysis
CSQalb.
CFS/serum quotient diagram according to Reiber and Lange (1991)
21
EUROIMMUN Microplate ELISA
EUROIMMUN
ELISA Automation Using the EUROIMMUN Analyzers

EUROIMMUN Analyzer I and EUROIMMUN Analyzer I-2P: Broad Parameter Spectrum, Proven Reliability, Variable Throughput
One for all: fully automated processing of all EUROIMMUN ELISA autoimmune diagnostics, infectious serology and allergology with only one system Flexibility for your benefit: open system with more than 800 validated EUROIMMUN parameters for serum, plasma and cerebrospinal fluid, over 50 or 30 parameters in parallel. Convenient, simple and reliable operation: e.g. by scanning QC certificates using a 2D-hand barcode scanner ready-for-use reagents and preprogrammed assay protocols enable you to start immediately. Capacity and throughput: quick loading and efficient time management allow processing of up to 70 or 50 tests per hour up to 7 or 3 plates, 180 or 144 samples at the start of a run. Security for patients: validated test systems and the comprehensive safety kit provide the basis for reliable diagnostics. Reliability and service: instruments and reagents from one manufacturer, quick and targeted support from our personnel for a smooth workflow in your laboratory.
Modular System: A Highly Sophisticated Solution

High convenience, fast and reliable loading through barcode identification of samples and reagents: automatic scanning when racks are inserted, reading of lot-specific QC data via 2D hand barcode scanner. Dilution area: 288 or 192 dilution positions (Deepwell, 2 ml). Liquid level detection (capacitive), multi-shot (dispensing) mode, automatic tip detection, clot detection. Pipetting possible during plate transport due to separation of transport and pipetting unit. 4 or 2 incubators with heating and shaking function, 4 or 3 incubators at room temperature. Standard Windows software: familiar user interface, all relevant statistical functions provided, available in different languages. Efficient use and convenient handling of tips and dilution plates through memory function.
A Convincing and Reliable Package: EUROIMMUN Analyzer, EUROIMMUN ELISA, EUROIMMUN Service
Comprehensive test system validation for the EUROIMMUN Analyzer: all parameters validated in accordance with the 98/79/EC directive and based on EN ISO 13485:2003/CMDCAS. All ELISA test systems are manufactured according to the European Quality Standards (IVD). National and international conformity (standardisation): CE, IVD, FDA and CMDCAS. Programming and set-up of automated system, including introduction to the system with customer training, performed by qualified personnel. Reliable and fast delivery of consumables and reagents. Connection to in-house computer system via communication protocol ASTM. Maintenance contract with EUROIMMUN on request.

ap
pr
ov e d
ap
22
pr
ov e d
*) soon available for the EUROIMMUN Analyzer I-2P
EUROIMMUN
microplate wells coated with antigens
Pipette:
100 l per well
;;; ;;; ;; ;; ;; ;; ;; ;;
diluted samples
Incubate:
30 min
Wash:
300 l wash buffer per well
3x
;; ;; ;; ;; ;; ;; ;; ;; ;; ;; ;; ;; ;;;; ;; ;;
;;; ;;; ;;; ;;; ;;; ;;; ;;; ;;; ;;; ;;; ;;; ;;; ;;; ;;; ;;; ;;; ;;; ;;; ;;; ;;; ;;; ;;; ;;; ;;;
enzyme conjugate
Pipette:
100 l per well
Incubate:
30 min
Wash:
3x
;; ;;;
stop solution chromogen/ substrate solution
Pipette:
100 l per well
Incubate:
15 min
;; ;; ;; ;; ;;;; yyy ;;; @@@ yyy @@@ ;;; yy @@ ;; ;; ;;;; ;; yy ;; @@ ;; ;; ;; @@ @@@ ;;; yyy @@@ ;;; yyy ;; ;; ;;;; ;;yy
yy @@ yyy @@@ ;;; @@@ ;; ;;; yyy yyy yyy yy ;;; ;;; ;; @@@ @@@ @@ ;; ;; ;; ;; ;; yyy yyy yy @@@ @@@ @@ ;;; ;;; yyy yyy yy ;;; ;;; ;; @@@ @@@ @@ ;; ;; ;; ;; ;;;;
23
Pipette:
100 l per well
Evaluate:
photometric measurement at a wavelength of 450 nm
Incubating the Microplate ELISA
EUROIMMUN
EUROASSAY: Line Blot in TITERPLANE Technique Format
membrane coated with antigen specific human antibody
AP AP-labelled anti-human antibody AP chromogen substrate
stain
nRNP/Sm Sm SS-A SS-B
EUROASSAY Anti-ENA ProfilePlus: detection of antibodies against SS-A and SS-B in a case of Sjgrens syndrome.
24
yyyyyy ;;;;;;
Scl-70 Jo-1 Control band

Membrane strips coated with thin parallel lines of several purified, biochemically characterized antigens are used as solid phase. The membrane strips are fixed as BIOCHIPs in the fields of microscope slides. If the sample is positive, specific antibodies in the diluted serum sample attach to the antigens coupled to the solid phase. In a second incubation step, the attached antibodies react with AP-labelled antihuman antibodies. In a third step, the bound antibodies are stained with a chromogen/substrate solution which is capable of promoting a color reaction. An intense dark band at the line of the corresponding antigen appears if the serum sample contains specific antibodies. The microscope slides are incubated using the TITERPLANE Technique: samples and reagents are applied to the reaction areas of a reagent tray. The slides are then placed into the recesses of the reagent tray, where all test strips of one slide come into contact with the liquids, and the individual reactions begin simultaneously. Depending on the spectrum of antigens used, it is possible to analyze several antibodies next to each other and simultaneously under identical conditions.
Easy Handling
Several serum samples can be analyzed simultaneously on one and the same slide. Total time for performing the EUROASSAY test is about 100 minutes. During the washing procedure, reagents for the next incubation step can be applied to reagent trays. All incubation steps proceed at room temperature. Shaking the slides together with the reagent tray on a circulatory shaker ensures the best possible sensitivity. Low reagent consumption. Only 50 l each of diluted serum and reagent are needed per test field. Reagents ready for use (wash buffer: concentrate).
Reliable and Simple Evaluation

Since results are evaluated visually, there is no investment required for photometers, etc. The antigen bands are located at exactly defined positions, which means that evaluation of the test is much simpler than for Westernblots. Correct completion of the individual incubation steps in each test field is indicated by staining of the control band. Positive and negative results can be easily and reliably differentiated from each other. The intensity of the antigen bands correlates with the antibody titer. The antigens used are highly pure, mostly isolated by affinity chromatography. The membrane strips do not contain any superfluous proteins which might cause unspecific positive results. The incubated microscope slides can be stored for long periods. Results can be easily documented.
EUROIMMUN
slide membrane strips reagent tray
Pipette:
50 l per field
;; ;; ;; ;;; ;; ;; ;; ;; ;;; ;; ;;; ;;;;

diluted samples
Incubate:
30 min shaking on a circulatory shaker (300 rpm)
;; ;; ;; ;; ;;; ;;; ;; ;;
wash buffer
Wash:
Pipette:
50 l per field
;;; ;;; ;;; ;;; ;;; ;;; ;;; ;;; ;;; ;;; ;;; ;;; ;;; ;;; ;;;
enzyme conjugate ;;; ;;; ;;; ;;; ;;; ;;; ;;; ;;; ;;; ;;;
Incubate:
30 min shaking on a circulatory shaker (300 rpm)
;; ;;;; ;;;; ;;;; ;;;; ;;

wash buffer
Wash:
Pipette:
50 l per field
Incubate:
10 min shaking on a circulatory shaker (300 rpm) flush with deionized or distilled water, air dry
;; ;; ;; ;; ;; ;; ;; ;; ;; ;; ;; ;; ;; ;; ;; ;; ;; ;; ;; ;; ;; ;; ;; ;; ;; ;; ;; ;; ;; ;;
chromogen/substrate solution
Wash:
Evaluate:
visual assessment of color intensity
25
Incubating the EUROASSAY (TITERPLANE Technique)
EUROIMMUN
The EUROLINE: A New Technique for Extensive Antibody Profiles

stain

substrate AP AP-labelled anti-human antibody AP chromogen
Membrane strips coated with thin parallel lines of several purified, biochemically characterized antigens are used as solid phase. The membranes are fixed as BIOCHIPs onto synthetic foil. If the sample is positive, specific antibodies in the diluted serum sample attach to the antigens coupled to the solid phase. In a second incubation step, the attached antibodies react with alkalinephosphatase-labelled anti-human antibodies. In a third step, the bound antibodies are stained with a chromogen/substrate solution which is capable of promoting a color reaction. An intense dark band at the line of the corresponding antigen appears if the serum sample contains specific antibodies. Depending on the spectrum of antigens used, it is possible to analyze several antibodies next to each other and simultaneously under identical conditions.
yyyyyy ;;;;;;
AMA M2 M2-3E (BPO) Sp100 nRNP/Sm Sm SSA Ro-52 SS-B
membrane coated with antigen
Easy Handling, Reliable and Simple Evaluation

Mi-2 Ku PM-Scl100 PM-Scl75 Scl-70 PML PM-Scl gp210 Jo-1 CENP B LKM-1 PCNA PL-12 LC-1 dsDNS Nucleos. EJ Histones SLA/LP Rib. P-prot. Ro-52 AMA-M2 OJ Ro-52 Jo-1 SRP PL-7 A separate membrane strip is incubated for each serum sample. Total time for performing the analysis is about 105 minutes. The incubation can be automated using the EUROBlotMaster. All incubation steps proceed at room temperature. The antigen bands are located at exactly defined positions, which means that the evaluation of the test is much simpler than for Westernblots. Correct completion of the individual incubation steps is indicated by staining of the control band on each EUROLINE test strip. Positive and negative results can be easily and reliable differentiated from each other. The intensity of the antigen bands correlates with the antibody titer. The antigens used are highly pure, mostly isolated by affinity chomatography. The membrane strips do not contain any superfluous proteins which might cause unspecific positive results. The incubated EUROLINE test strips can be stored for long periods. Results can be easily documented. The program EUROLineScan from EUROIMMUN has been developed to enable quantitative evaluation of EUROLINE test strips, to facilitate management of data, and to provide detailed documentation of results. First, the incubated EUROLINE test strips are scanned using a flatbed scanner. EUROLineScan recognizes the position of the strips, even if they have been placed inexactly, identifies the bands, and measures their intensity. The results are then saved together with the image data. A separate results sheet can be produced for each patient.
Control
Control
Control
Incubated EUROLINE test strips (Autoimmune Liver Diseases Profile, ANA Profile 3, Myositis Profile 3.
26
EUROIMMUN
EUROBlotMaster
Standardised incubation of immunoblot strips higher precision and reproducibility. Automatisation of all EUROIMMUN immunoblot strips (EUROLINE, EUROLINEWB, Westernblot) Over 65 validated parameters available (16 autoantibody, 28 infectious and 21 allergy parameters) Up to 30 strips per test run Easy operation Combination of different conjugates/tests in one run Walk-away function fully automated from the start to the end of processing following loading Compatible with modern evaluation systems such as EUROBlotCamera and EUROLineScan EUROBlotMaster
Automatic Evaluation of the Results Using EUROLineScan

For all EUROIMMUN blot systems: EUROLINE, EUROLINE-WB, Westernblot. For all areas: autoimmune diagnostics, infectious serology and allergology. EUROBlotCamera: digitalisation of strips while in the incubation tray. EUROBlotScanner: digitalisation of strips using flatbed scanner. Fully automated identification, quantitation and assignment of bands. Option to modify results (changes are automatically documented). Complete results obtained just a few minutes after finishing the incubation. Fully automated administration and documentation of extensive individual data. Electronic archiving of all images and data (avoids the need to store potentially infectious blot strips). Online connection to laboratory software. Network compatible.
EUROBlotCamera
EUROBlotScanner
27
EUROLINE Automation Using EUROBlotMaster and EUROLineScan
EUROIMMUN
Westernblot/EUROLINE-WB: Reliable Differentiation of Antibodies Present

stain chromogen AP AP-labelled anti-human antibody AP

Membrane strips containing electrophoretically separated antigen extracts are used as solid phase. The position of the proteins depends on their respective molecular masses. If the sample is positive, specific antibodies in the diluted serum sample attach to the antigens coupled to the membrane. In a second incubation step, the attached antibodies react with AP-labelled antihuman antibodies. In a third step, the bound antibodies are stained with a chromogen/substrate solution which is capable of promoting a color reaction. An intense dark band at the line of the corresponding antigen appears if the serum sample contains specific antibodies. Evaluating the band patterns on the incubated membrane strips involves differentiating non-specific from specific antibodies. The number and intensity of the specific bands is decisive for the result positive/negative.
substrate
yyyyyy ;;;;;;
membrane coated with antigen
VlsE antigen on EUROLINE membrane chip
Easy Handling, Reliable Evaluation and High Diagnostic Significance

A separate membrane strip is incubated for each serum sample. Total time for performing the Westernblot test is about 115 minutes. All incubation steps proceed at room temperature. The bands are assigned according to a lot-specific evaluation matrix provided. A separate lot is issued for each electrophoresis gel, helping to avoid errors in the assignment of the bands. Every test kit contains a membrane strip of the same lot incubated with a positive reference serum. Therefore, there is no need to incubate a positive control serum. The membrane strips are pre-numbered to prevent confusion. Laborious labelling is not necessary. Correct completion of the individual incubation steps for each membrane strip is indicated by staining of the control band at the bottom of the strip. Positive and negative reactions can be easily and reliably differentiated from each other. The intensity of the antigen bands correlates with the antibody titer. The Westernblot is the method of choice when the objective is to confirm or differentiate positive results obtained in a screening test (indirect immunofluorescence or microplate ELISA). EUROLINE-WB is a combination of westernblot and line blot techniques. Proteins from a whole antigen extract are electrophoretically separated according to molecular mass and transferred onto a nitrocellulose membrane. Highly purified native or recombinant antigens are then printed as lines onto the westernblot strips (EUROLINE membrane chip). The program EUROLineScan from EUROIMMUN has been developed to enable quantitative evaluation of Westernblot/EUROLINE-WB test strips, to facilitate management of data, and to provide detailed documentation of results. First, the incubated Westernblot/EUROLINE-WB test strips are scanned using a flatbed scanner. EUROLineScan recognizes the position of the strips, even if they have been placed inexactly, identifies the bands, and measures their intensity. The results are then saved together with the image data. A separate results sheet can be produced for each patient.
p 83
p 41, Flag. p 39, Bmp A p 31, Osp A p 30
p 25, Osp C p 21 p 19 p 17 Alignment bar

EUROLINE-WB Borrelia
Control band
EUROLINE-WB: detection of antibodies against Borrelia.
28
EUROIMMUN
using the EUROBlotMaster or manually on a rocking platform
membrane strip incubation channel buffer Pipette: 1.5 ml per channel
Incubate:
5-15 min shaking, depending on test system
Aspirate off:
Pipette:
1.5 ml per channel
;;; ;;;
diluted samples
Incubate:
30 min shaking
;;;;;;;;;;;;;;;;;; ;;;;;;;;;;;;;;;;;;
Wash:
1.5 ml buffer, 5 min incubation, aspirate off
3x
;;; ;;;
buffer
enzyme conjugate
Pipette:
1.5 ml per channel
Incubate:
30 min shaking
;;;;;;;;;;;;;;;;;;;;;;;;;;;; ;;;;;;;;;;;;;;;;;;;;;;;;;;;; ;;;;;;;;;;;;;;;;;;;;;;;;;;;;
Wash:
1.5 ml buffer, 5 min incubation, aspirate off
3x
buffer
Pipette:
1.5 ml per channel
Incubate:
10 min shaking
;; ;; ;;;;;;;;;;;;;;; ;;;;;;;;;;;;;;;
chromogen/substrate solution
Wash:
rinse with distilled water, aspirate off with EUROLineScan or visual assessment
Evaluate:
Applicable for most EUROLINE/Westernblot/EUROLINE-WB test kits
29
Incubating the EUROLINE/Westernblot/EUROLINE-WB
EUROIMMUN
EUROIMMUN Radioimmunoassays (RIA/IRMA)

Test principle RIA (precipitation techniques)
radioactively labelled antigen (tracer)
125
125
In the first incubation step patient sera are incubated with 125I-labelled antigen in polystyrene tubes. Any specific antibodies in the sera bind to the antigen. In the second incubation step the antigen-antibody complexes are precipitated using a precipitation agent. The precipitate is washed with buffer. After centrifugation and decanting of the supernatant, the radioactivity in the precipitate is counted using a gamma counter. The intensity of the radioactivity is proportional to the concentration of specific antibody in the patient serum. The antibody concentration is evaluated quantitatively using a calibration curve.
precipitation agent
Test principle RIA (coated tubes)

RIA tests (coated tubes) are competitive ligand assays for antibody and antigen determinations. The intensity of radioactive radiation is inversely proportional to the concentration of specific antibodies or antigens in the sample. The quantitative evaluation of the antigen/antibody concentration is carried out using a calibration curve.
Test principle IRMA (antigen determination)

solid phase (tube surface)
radioactively labelled antibody

125
analyte
antibody
With this test principle, monoclonal antibodies which are bound directly or indirectly to the inner wall of polystyrene tubes are used. During the first incubation step, the patient sera to be investigated are incubated with the monoclonal 125I-labelled antibodies in the coated tubes. The antigen in the sample is bound by the immobilised antibodies and by the 125 I-labelled antibodies. This results in a solid-phase bound sandwich complex. The unbound 125I-labelled antibodies are removed by washing and subsequently decanting. The intensity of radioactive radiation is proportional to the concentration of antigens in the patient serum. The quantitative evaluation of the antigen concentration is carried out using a calibration curve.
Simple and economical handling, reliable analysis

Simple test procedure. Synchronous processing of samples, including different tests in parallel. Ready-to-use reagents (possible exceptions: tracer, precipitation reagents). Different test formats for small and large sample series. Because of the large measurement range of EUROIMMUN RIA, further measurements with other sample dilutions are generally not necessary. Simple evaluation of test results. Each test can be optionally evaluated with controls which are supplied in the test kit. Test kit-specific reference ranges are given for all controls. EUROIMMUN radioimmunoassays show the following analytical characteristics: High analytical specificity. High detection sensitivity. High clinical sensitivity and specificity. Good reproducibility.
30
EUROIMMUN
EUROIMMUN PRODUCTS FOR THE DETERMINATION OF AUTOANTIBODIES
31
EUROIMMUN Products for the Determination of Autoantibodies
EUROIMMUN
Autoantibodies against Cell Nuclei (ANA)

Indirect Immunofluorescence Test: EUROPLUS ANA Mosaic 20
Screening test for the detection of antibodies against cell nuclei (ANA). Indications: rheumatic diseases. Initial dilution 1 : 100; conjugate class anti-human IgG, FITC-labelled. Using HEp-2 cells many antibodies against cell nuclei can be analyzed, e.g. antibodies against DNA, histones, RNA, nRNP, Sm, SS-A, SS-B, nuclear dots, centromeres, nuclear membrane, nucleoli (PM-Scl, fibrillarin, RNA polymerase I, NOR), Scl-70, cyclin I and II, spindle fibers, midbody, centrioles. In addition, cytoplasmic autoantibodies are identified with HEp-2 cells: antibodies against ribosomes, Jo-1, mitochondria, Golgi apparatus, actin etc. The primate liver permits the verification of results between both substrates, makes the pre-differentiation of a large number of ANA possible, and helps to establish titer levels. Moreover, the primate liver often contains additional antigens, allowing the identification of further autoantibodies: antibodies against LMA, LSP, endomysium, bile ducts and endothelium cells, as well as cANCA und pANCA. The EUROPLUS system is a monospecific test that can be used to confirm the presence of autoantibodies against cell nuclei and cytoplasm with one and the same test kit. The following antigens are currently available as EUROPLUS BIOCHIPs: SS-A, SS-B, nRNP/Sm, Sm, Scl-70, Jo-1, ribosomal P-proteins.
EUROPLUS ANA Mosaic 20 (HEp-2 cells, primate liver, SS-A+SS-B BIOCHIPs, rib. P-prot.+Jo-1 BIOCHIPs: antibodies against SS-A/SS-B. Order No. FA 1510-####-20 G Formats page 126
Indirect Immunfluorescence Test: Innovative Cell Line from EUROIMMUN, HEp-20-10

Screening test for detection of antibodies against cell nuclei. Indications: rheumatic diseases. Initial dilution 1 : 100; conjugate class anti-human IgG, FITC-labelled. Compared to standard HEp-2 cells, HEp-20-10 cells demonstrate 10-fold more mitotic cells. Antibodies against mitotic-specific structures (centromeres, spindle fibers, midbody, centrioles, NOR) can be more easily identified than with conventional preparations. At the same time, the cell line HEp-20-10 offers the full antigen spectrum for cell nuclei antibody diagnostics. The BIOCHIP with HEp-20-10 can be supplemented with additional substrates, for example, primate liver (ANA; Order No. FA 1512-####-1 G) as well as rat kidney and rat stomach (Order No. FA 1802-####-3 G).
HEp-2010: antibodies against spindle fibers.
Order No. FA 1522-#### G
Formats page 128
EUROASSAY: Anti-ENA ProfilePlus

Differentiation of anti-nuclear antibodies (ANA). Indications: rheumatic diseases. Serum dilution 1 : 100; conjugate class anti-human IgG, AP-labelled. 6 relevant anti-nuclear antibodies against extractable nuclear antigens (ENA) can be detected simultaneously and monospecifically: antibodies against nRNP/Sm, Sm, SS-A, SS-B, Scl-70, Jo-1. Native antigens, purified by affinity chromatography. On request EUROASSAY can be produced with special antigen combinations, for example, with dsDNA, histones, nucleosomes, PM-Scl, nRNP/Sm, Sm, SS-A, Ro-52, SS-B, Scl-70, Jo-1, ribosomal P proteins, centromere protein B, M2, M4, M9, SLA/LP, LC-1, LKM-1.
Incubated EUROASSAY Anti-ENA ProfilePlus.
Order No. DA 1590-####-1 G
Formats page 79
32
EUROIMMUN

EUROLINE: ANA Profile 3

Differentiation of antibodies against cell nuclei (ANA). Indications: Sharp syndrome (MCTD), systemic lupus erythematosus, Sjgrens syndrome, progressive systemic sclerosis, poly/dermatomyositis, PBC. Serum dilution 1 : 100; conjugate class anti-human IgG, AP-labelled. With the EUROLINE ANA-Profile 3, fifteen autoantibodies can be determined: antibodies against nRNP/Sm, Sm, SS-A, Ro-52, SS-B, Scl-70, PM-Scl, Jo-1, centromere protein B, PCNA, dsDNA, nucleosomes, histones, ribosomal Pproteins, AMA M2. Antibodies against SS-A are characteristic markers for SLE and Sjgrens syndrome. In contrast, antibodies against Ro-52 also occur in patients with other autoimmune diseases. Native antigens, purified by affinity chromatography (exception: centromere protein B, PM-Scl, Ro-52, PCNA). Further antigen combinations: page 80. Test strips can be automatically incubated and evaluated using the systems EUROBlotMaster und EUROLineScan (see page 27).
nRNP/Sm Sm SS-A Ro-52 SS-B Scl-70 PM-Scl Jo-1 CENP B PCNA dsDNA nucleos. histones rib. P-prot. AMA M2 control
Incubated EUROLINE ANA Profile 3.
Order No. DL 1590-1601-3 G
Formats page 80
EUROLINE: Myositis Profile 3

Differentiation of myositis-associated antibodies against cell-nuclear and cytoplasmic antigens. Indications: poly/dermatomyositis. Serum dilution 1 : 100, conjugate class anti-human IgG, AP-labelled. With the EUROLINE Myositis Profile 3, eleven autoantibodies can be determined: antibodies against Mi-2, Ku, PM-Scl100, PM-Scl75, Jo-1, SRP, PL-7, PL-12, EJ, OJ and Ro-52. Test strips can be automatically incubated and evaluated using the systems EUROBlotMaster und EUROLineScan (see page 27).
Mi-2 Ku PM-Scl100 PM-Scl75 Jo-1 SRP PL-7 PL-12 EJ OJ Ro-52 control
Incubated EUROLINE Myositis Profile 3.
Order No. DL 1530-1601-3 G
Formats page 80
EUROLINE-WB: HEp-2 Cell Antigens plus SS-A, Ro-52 and CENP B

Differentiation of antibodies against cell-nuclear and cytoplasmic antigens. Indications: Sharp syndrome (MCTD), systemic lupus erythematosus, Sjgrens syndrome, progressive systemic sclerosis, poly/dermatomyositis, PBC. EUROLINE-WB is a combination of westernblot and line blot techniques. Proteins from a whole antigen extract of HEp-2 cells are electrophoretically separated according to molecular mass and transferred onto a nitrocellulose membrane. A membrane chip coated with highly purified native SS-A, recombinant Ro-52 and recombinant CENP B is then added to the westernblot strips. Antibodies against SS-A are characteristic markers for SLE and Sjgrens syndrome. In contrast, antibodies against Ro-52 also occur in patients with other autoimmune diseases. EUROLINE-WB contains both antigens next to each other at defined positions, in addition to the complete HEp-2 antigen spectrum of the Westernblot. The use of native SS-A increases the sensitivity, since 37% of antibodies against SS-A do not show any reaction with the denatured antigen from the Westernblot Serum dilution 1 : 50, conjugate class anti-human IgG, AP-labelled. Antigens: SDS extract from HEp-2 cells (whole antigen), highly purified native SS-A from calf thymus, recombinant Ro-52 and CENP B.
rec. CENP B CENP B Ku RNP 70
native SS-A rec. Ro-52
SS-A Ro-52
RNP A Sm B/B
Rib.-P
Incubated EUROLINE-WB HEp-2 Cell Antigens plus SS-A, Ro-52 and CENP B. Order No. DW 1520-1601-3 G Formats page 83
33
EUROIMMUN

Microplate ELISA: ANA Screen, Anti-ENA PoolPlus
Incubated ELISA ANA Screen (antigen mixture of dsDNA, histones, nRNP/Sm, Sm, SS-A, SS-B, Scl70, Jo-1, ribosomal P-proteins, centromere). Screening test for predifferentiation of antibodies against cell nuclei (ANA) and cytoplasm components. Indications: Sharp syndrome (MCTD), systemic lupus erythematosus, Sjgrens syndrome, progressive systemic sclerosis, polymyositis/dermatomyositis. Serum dilution 1 : 200, conjugate class anti-human IgG, POD-labelled. One microplate well incubated per patient. 1-point calibration, semiquantitative. Native antigens (exception: centromere, recombinant). The ANA Screen ELISA supplements the gold standard immunofluorescence. It is based on a mixture of 10 highly purified antigens, which provide higher sensitivity and specificity than the undefined cell extracts used by other manufacturers. Two ELISAs with different antigen combinations, adapted to particular indications or for follow-up of immunofluorescence patterns, are available.
Order No. EA 1590-9601-8 G
Formats page 87
Microplate ELISA: SLE Profile 1/2, Anti-ENA ProfilePlus 1/2

Differentiation of antibodies against cell nuclei (ANA) and cytoplasm components. Indications: Sharp syndrome (MCTD), systemic lupus erythematosus, Sjgrens syndrome, progressive systemic sclerosis, polymyositis/dermatomyositis. Serum dilution 1 : 200; conjugate class anti-human IgG, POD-labelled. 8 or 6 relevant antibodies can be detected simultaneously. 1-point calibration, semiquantitive. Calibrator pool and negative controls each on a separate microplate strip (SLE Profiles and Anti-ENA ProfilePlus 2) or on the same microplate strip as the patient serum. Native antigens (exception: Ro-52, centromere and PM-Scl, recombinant). In total 4 different ELISAs with different antigen combinations, adapted to particular indications or for follow-up of immunofluorescence patterns, are available.
Incubated ELISA Anti-ENA ProfilePlus 2 (antigens: ribosomal P-proteins, nRNP/Sm, Sm, SS-A, SS-B, Scl-70, Jo-1, centromere).
Order No. EA 1590-1208-2 G
Formats page 87
Microplate ELISA: ANA Single-Antigen ELISAs

Incubated ELISA Anti-SS-A, Anti-SS-B. Differentiation of antibodies against cell nuclei (ANA) and cytoplasm components. Indications: rheumatic diseases. Serum dilution 1 : 200; conjugate class anti-human IgG, POD-labelled. Antibodies against cell nuclei components can be determined quantitatively in RU/ml. 3-point calibration, quantitative. Identical incubation conditions and times: all single-antigen tests can be combined with each other on one microplate. Native antigens (exceptions: centromere and PM-Scl, recombinant). Single-antigen ELISAs available for detection of antibodies against cell nuclei and cytoplasm antigens: ssDNA, nucleosomes, dsDNA, histones, ribosomal P proteins, PM-Scl, nRNP/Sm, Sm, SS-A, SS-B, Scl-70, Jo-1, centromere.
Order No. EA ####-9601 G
Formats page 87
34
EUROIMMUN
Autoantibodies against Double-Stranded DNA (dsDNA)

Indirect Immunofluorescence Test: Crithidia luciliae

Detection of antibodies against dsDNA. Indication: lupus erythematosus disseminatus. Initial dilution: 1:10, conjugate class anti-human IgG, FITC-labelled. Animal pathogenic haemoflagellates of Crithidia luciliae are used for the detection of autoantibodies against double-stranded, native DNA (dsDNA, nDNA) with indirect immunofluorescence. These protozoa possess a giant mitochondrion containing dsDNA (kinetoplast) that, in general, does not show any of the other antigens present in the cell nuclei. Antibodies reacting with the kinetoplast are therefore only directed against dsDNA. Alongside the conventional CLIFT, which shows a particularly high disease specificity, EUROIMMUN has developed an Anti-Crithidia luciliae sensitive IFT (order no. FA 1572-####-1), which is comparable in sensitivity to the Anti-dsDNANcX ELISA and the Farr assay. However, despite comparable sensitivities, the assays identify different patients. To increase the serological hit rate different test systems are often combined.
Crithidia luciliae: antibodies against dsDNA.
Order No. FA 1572-####
Formats page 128
Microplate ELISA: Anti-dsDNA-NcX

Monospecific detection of antibodies against dsDNA. Indication: lupus erythematosus disseminatus. Serum dilution 1: 200, conjugate class anti-human IgG, POD-labelled. Antibodies against dsDNA can be determined quantitatively in IU/ml. 3-point calibration, quantitative. Antigen: double-stranded DNA, complexed with nucleosomes (NcX). Due to good sensitivity and specificity, the Anti-dsDNA-NcX ELISA stands out by high diagnostic efficiency. High concentrations of autoantibodies against dsDNA in the ELISA are considered to be a reliable marker for the diagnosis or prognosis of SLE. Individual changes in the dsDNA antibody concentration correlate with the activity of the disease and can be used for monitoring the development of the disease in SLE patients. In cases of immunosuppressive therapy or clinical remission dsDNA antibodies cannot be detected with the ELISA anymore.
Incubated ELISA Anti-dsDNA-NcX.
Order No. EA 1572-9601 G
Formats page 87
Anti-dsDNA RIA by Farr

Monospecific detection of antibodies against dsDNA. ndication: lupus erythematosus disseminatus. Use of undiluted samples. Antigen: 125I-labelled dsDNA from plasmid DNA. The Farr radioimmunoassay has always been of great importance. On the whole, it has the same specificity as the immunofluorescence test and the same sensitivity as the ELISA. Apparently, its well-known high diagnostic efficiency is based on the fact that only the fraction of the anti-dsDNA antibodies which is able to form bigger precipitating immune complexes with circulating DNA in liquid phase contributes to the measuring signal. The principle of the Farr test reflects, so to speak, the significant step in the pathomechanism of SLE: the formation of appropriate immune complexes, deposits of which build up in the joints, kidneys, liver and other organs.
RIA Anti-dsDNA.
Order No. RA 1571-0001
Formats page 89
35
EUROIMMUN
Autoantibodies against CCP and Sa

Microplate ELISA: Anti-CCP, Anti-Sa
Screening test for the specific determination of autoantibodies against cyclic citrullinated peptides (CCP) and citrullinated Sa. Indication: rheumatoid arthritis (RA). Serum dilution 1 : 100; conjugate class anti-human IgG, POD-labelled. Antibodies against CCP and Sa can be determined quantitatively in RU/ml. Optional reference control for the determination of ratio values. Antigen: synthetic cyclic citrullinated peptides (CCP, second generation), citrullinated Sa.
Antigen
CCP Incubated ELISA Anti-CCP, Anti-Sa. Sa Order No. EA 1505-9601 G EA 151a-4802 G
Order No. (Anti-CCP) EA 1505-9601 G
Formats page 87
Antibodies against cyclic citrullinated peptides (CCP): An ELISA for specic diagnosis of rheumatoid arthritis
H
colourless chromogen
stain
H
N
Peptidylargininedeiminase (PAD) Ca2+
O
POD
peroxidase-labelled anti-human antibody

POD
NH H 2N+
NH O
human antibody against CCP
NH2
NH2
synthetic CCP
L-arginine
L-citrulline
The amino acid citrulline
Principle of the anti-CCP ELISA
Anti-CCP ELISA
Rheumatoid arthritis (RA) is one of the most common autoimmune diseases, affecting around 1% of the world population. It is characterised by inammation of the synovial membrane, which spreads symmetrically from the small to the large joints. Initial symptoms include painful swelling of nger joints with morning stiffness in the joints. Early diagnosis and immediate commencement of suitable therapy is necessary to keep the disease under control. The most commonly performed serological test in suspected RA cases was until now the determination of rheumatoid factors (RF). These are antibodies (predominantly of class IgM) which react with gamma globulins and occur in 60-80% of RA patients. RF are a sensitive but not very specic marker for RA, since they also occur in healthy individuals and in patients with various infections or other autoimmune diseases (systemic lupus erythematosus, Sjgrens syndrome, scleroderma and others). 40-60% of RA patients also exhibit autoantibodies against epidermal laggrin1 (RA keratin, anti-perinuclear factor) in their serum. Filaggrin is a protein of the epidermis, which links keratin laments to one another. Autoantibodies against laggrin are detected by indirect immunouorescence: the antigen substrate rat oesophagus shows staining of the stratum corneum (RA keratin) on the luminal side; anti-perinuclear factors (APF) are apparent in the cytoplasmic inclusion bodies of human epithelial cells of the oral mucosa.
In recent years it has been shown that the rare amino acid citrulline, which is present in laggrin, is a substantial component of the antigenic epitope. Enzyme immunoassays which use synthetic citrullinated peptide as the target antigen offer a useful alternative to indirect immunouorescence2. A direct comparison study demonstrated that the sensitivity can be increased from 49% to 68% by using cyclic citrullinated peptide instead of linear citrullinated peptide as an ELISA substrate3. Antibodies against cyclic citrullinated peptides (CCP) are a new and highly specic marker for RA. Antibodies against CCP are predominantly of class IgG and have a specicity of 98% for RA. They are observed very early in the disease course and have a high predictive value: patients with anti-CCP antibodies develop signicantly more radiologically detectable joint damage than anti-CCPnegative patients4. Antibodies against CCP possess a much higher specicity than RF (anti-CCP: 97%, RF: 62%) with the same sensitivity (anti-CCP: 79%, RF: 78%)5. They can be detected in early stages of the disease in 79% of patients. EUROIMMUN offers an innovative microplate ELISA for quantitative determination of autoantibodies against CCP. Diluted patient sera are incubated in wells coated with synthetic cyclic citrullinated peptides (second generation). Specic antibodies in the serum bind to the immobilised antigen and cause a photometric colour reaction by means of an enzyme-coupled secondary antibody. Five
calibration sera ensure reliable measurement of antibody concentrations. The EUROIMMUN Anti-CCP ELISA is a highly specic and sensitive serological test system for the diagnosis of RA.
Panel Sensitivity RA Asymptomatic blood donors Psoriatic arthritis Other arthritides Systemic lupus erythematosus Sjgrens syndrome Scleroderma Autoimmune thyroiditis Wegener granulomatosis Anti-parvovirus B19 positive Viral hepatides Anti-HIV positive Tuberculosis Specicity RA n 419 400 28 35 108 106 98 159 25 126 54 5 10 1154 Anti-CCP positive 329 (78.5%) 2 (0.5%) 0 3 (8.6%) 3 (2.8%) 2 (1.9%) 3 (3.1%) 4 (2.5%) 1 (4.0%) 3 (2.4%) 0 0 0 21 (98.2%)
1) Nogueira et al., Ann. Rheum. Dis. 60: 882 (2001) 2) Schellekens et al., J. Clin. Invest. 101: 273-281 (1998) 3) Schellekens et al., Arthritis Rheum. 43: 155-163 (2000) 4) Kroot et al., Arthritis Rheum. 43: 1831-1835 (2000) 5) Vasishta, Am. Clin. Lab. 21: 34-36 (2002)
36
EUROIMMUN
Autoantibodies against Mitochondria (AMA)

Indirect Immunofluorescence Test: EUROPLUS Rat Kidney and M2-3E BIOCHIPs

Screening test for the detection of antibodies against mitochondria (AMA) including simultaneous confirmation of the subtype AMA M2. Indication: primary biliary cirrhosis (PBC). Initial dilution 1 : 100; polyvalent conjugate anti-human IgAGM, FITC-labelled. Rat kidney is the standard substrate for detecting anti-mitochondrial antibodies. Nine AMA types (M1 to M9) can be differentiated. The BIOCHIP coated with M2-3E permits monospecific confirmation of antibodies against the native pyruvate dehydrogenase complex and the recombinant M2 fusion protein (BPO) in one single test procedure, thus a PBC can be diagnosed serologically with confidence. This BIOCHIP Mosaic can be supplemented as required using additional substrates, e.g. HEp-2 cells (ANA, nuclear dots), rat liver (liver-kidney microsomes, LKM) or rat stomach (ASMA).
Rat kidney and M2-3E BIOCHIP: antibodies against mitochondria (AMA).
Order No. FA 1620-####-3
Formats page 129
EUROASSAY: AMA Profile M2, M4, M9

Differentiation of mitochondrial antibodies (AMA). Indication: primary biliary cirrhosis (PBC). Serum dilution 1 : 100; conjugate class anti-human IgGM, AP-labelled. 3 relevant mitochondrial antibodies can be detected simultaneously and monospecifically: antibodies against M2, M4, M9. Native antigens: pyruvate dehydrogenase complex (M2), sulfite oxidase (M4), glycogen phosphorylase (M9). AntiM2 M4 M9 Associated diseases Primary biliary cirrhosis (high titers), other chronic liver diseases Primary biliary cirrhosis Early phase of primary biliary cirrhosis
Incubated EUROASSAY AMA Profile M2, M4, M9.
Order No. DA 1620-####-1 O
Formats page 80
Microplate ELISA: Anti-M2-3E

Differentiation of mitochondrial antibodies (AMA). Indication: primary biliary cirrhosis (PBC). Serum dilution 1 : 100; conjugate class anti-human IgG, POD-labelled. Antibodies against M2 can be determined quantitatively in RU/ml. Antigen: native pyruvate dehydrogenase complex plus recombinant M2 fusion protein (BPO) containing the immunogenic domains of the E2 subunits of PDH, BCOADH and OGDH.
Incubated ELISA anti-M2-3E.
Formats page 88
37
EUROIMMUN
Autoantibodies against Liver Antigens

Indirect Immunofluorescence Test: Liver Mosaic 8
Screening and differentiation test for the detection of liver-specific antibodies, antibodies against mitochondria (AMA), antibodies against cell nuclei (ANA), antibodies against smooth muscles (ASMA), F-actin and other autoantibodies. Indications: autoimmune hepatitis, primary biliary cirrhosis, rheumatic diseases. Initial dilution 1 : 100; polyvalent conjugate anti-human IgAGM, FITC-labelled. The BIOCHIP Mosaic consists of 6 substrates: human epithelial cells (HEp-2), primate liver, rat kidney, rat liver, rat stomach, VSM47. Thus, a broad spectrum of antigens is present, allowing not only targeted serological diagnoses, but also frequently yielding additional results with clinical relevance. Antibodies against cell nuclei (ANA) can be particularly well demonstrated using HEp-2 cells and primate liver, and are identified according to their fluorescence patterns. However, they also stain the cell nuclei of the other tissues more or less intensely. Clinical significance: rheumatic diseases, primary biliary cirrhosis (antibodies against nuclear dots). With primate liver, several liver-specific autoantibodies can be investigated e.g. antibodies against liver cell membrane (anti-LMA) and liver-specific protein (anti-LSP). Clinical significance: autoimmune hepatitis. Antibodies against mitochondria (AMA) show a granular cytoplasmic fluorescence on all 6 substrates. With the standard substrate rat kidney, the proximal and distal tubule cells fluoresce equally. Clinical significance: primary biliary cirrhosis. Autoantibodies against liver-kidney microsomes (anti-LKM) react with rat liver and rat kidney (see below). The other substrates are essentially negative. In the case of autoantibodies against smooth muscles (ASMA), the tunica muscularis, the lamina muscularis mucosa as well as the interglandular contractile fibrils fluoresce on the rat stomach. ASMA directed against the target antigen F-actin furthermore react with the cytoskeleton of HEp-2 cells and the bile canaliculi of primate liver. The substrate VSM47 reacts very specifically, showing a filamentous, needle-like fluorescence. Clinical significance: autoimmune (lupoid) chronic active hepatitis. The BIOCHIP Mosaic can be supplemented as required with additional substrates, e.g. Crithidia luciliae (antibodies against dsDNA), musculus iliopsoas (antibodies against skeletal muscles), heart (antibodies against striated muscles, antibodies against intercalated disks, AMA M7), different EUROPLUS substrates (AMA-M2-3E, Sp100, gp210, PML, SLA/LP, LC-1, LKM).
HEp-2 cells: anti-nuclear dots. Primate liver: antiLSP. Rat kidney: AMA. Rat liver: ANA. Rat stomach: ASMA. VSM47: Anti-actin. Order No. FA 1300-####-8 Formats page 122
Indirect Immunofluorescence Test: BIOCHIP Mosaic Rat Liver/ Rat Kidney (Liver Mosaic 1)
Specific detection of antibodies against liver-kidney microsomes (anti-LKM). Indications: autoimmune hepatitis, often associated with extrahepatic syndromes such as arthralgias, glomerulonephritis, vitiligo and chronic inflammatory bowel diseases. Initial dilution 1 : 100; polyvalent conjugate anti-human IgAGM, FITC-labelled. Autoantibodies against liver-kidney microsomes react with rat liver and generate a smooth staining in the cytoplasm of the hepatocytes. In rat kidney, particularly in the cortex area, a fine granular fluorescence of the proximal tubules recognizable by the luminal brush border is visible. The distal tubules are negative. The fluorescence intensity of the liver cells is normally stronger than that of the proximal renal tubules. The differentiation between autoimmune hepatitis and virus-induced hepatitis can additionally be accomplished by investigating the appropriate viral parameters.
Rat liver and rat kidney: antibodies against liverkidney microsomes (anti-LKM).
Order No. FA 1300-####-1
Formats page 122
38
EUROIMMUN
Autoantibodies against Liver Antigens

EUROLINE: Profile Autoimmune Liver Diseases

Differentiation of antibodies in autoimmune liver diseases. Indications: autoimmune hepatitis, primary biliary cirrhosis, overlap syndromes. Serum dilution 1 : 100, conjugate class anti-human IgG, AP-labelled. With the EUROLINE Profile Autoimmune Liver Diseases, nine autoantibodies can be determined: antibodies against AMA M2, M2-3E (BPO), Sp100, PML, gp210, LKM-1, LC-1, SLA/LP and Ro-52. Test strips can be automatically incubated and evaluated using the systems EUROBlotMaster und EUROLineScan (see page 27). Further antigen combinations on page 79. AntiM2, Sp100, PML, gp210 LKM-1, SLA/LP, LC-1 Ro-52 Associated Diseases Primary biliary cirrhosis Autoimmune hepatitis Autoimmune hepatitis, rheumatic diseases Order No. DL 1300-1601-4 G Formats page 80
AMA M2 3E (BPO) Sp100 PML gp210 LKM-1 LC-1 SLA/LP Ro-52
control
Incubated EUROLINE Profile Autoimmune Liver Diseases.
EUROASSAY: Liver Profile (Anti-M2, -LKM-1, LC-1, -SLA/LP)

Determination of mitochondrial antibodies AMA M2, antibodies against liverkidney microsomes type 1 (LKM-1), antibodies against liver cytosolic antigen type 1 (LC-1), as well as of antibodies against soluble liver antigen/liver-pancreas antigen (SLA/LP). Indication: autoimmune liver diseases. Serum dilution 1 : 100, conjugate class anti-human IgG, AP-labelled. Antibodies against M2, LKM-1, LC-1 and SLA/LP can be detected simultaneously and monospecifically. Antigens: pyruvate dehydrogenase complex (M2, native), cytochrome P450 IID6 (LKM-1, recombinant), formiminotransferase-cyclodeaminase (LC-1, recombinant) and soluble liver antigen/liver-pancreas antigen (SLA/LP, recombinant).
Incubated EUROASSAY M2, LKM-1, LC-1, SLA/LP.
Order No. DA 1300-1003-3 G
Formats page 79
Microplate ELISA: Anti-SLA/LP, Anti-LC-1, Anti-LKM-1

Monospecific determination of antibodies against soluble liver antigen/liverpancreas antigen (SLA/LP), cytosolic liver antigen type 1 (LC-1) and liver-kidney microsomes type 1 (LKM-1). Indication: autoimmune hepatitis. Serum dilution 1 : 100, conjugate class anti-human IgG, POD-labelled. 3-point calibration, quantitative (exception: Anti-LC-1, semi-quantitative). Identical incubation conditions and times: all tests can be combined without difficulty on one and the same microplate. Recombinant antigens: soluble liver antigen/liver-pancreas antigen (SLA/LP), formiminotransferase-cyclodeaminase (LC-1) and cytochrome P450 IID6 (LKM-1). The corresponding human cDNA was expressed in E. coli (SLA/LP) or insect cells (LC-1, LKM-1).
Incubated ELISA Anti-SLA/LP, Anti-LC-1, Anti-LKM-1.
Order No. (Anti-SLA/LP) EA 1302-9601 G
Formats page 86
39
EUROIMMUN
Autoantibodies against Thyroid Gland Antigens / Antigen Detections

Indirect Immunofluorescence Test: EUROPLUS Thyroid Gland (unfixed) and Thyroglobulin
Detection of antibodies against thyroid gland antigens. Indications: Basedows disease, Hashimoto autoimmune thyroiditis. Initial dilution 1 : 10; conjugate class anti-human IgG, FITC-labelled. Using unfixed thyroid tissue, two important thyroid-specific antibodies can be found: Autoantibodies against thyroid microsomes (MAb) give a granular staining in the cytoplasm of the follicle epithelium (target antigen: thyroid peroxidase, TPO). Autoantibodies against thyroglobulin (TAb) react with the colloid of all follicles of the thyroid tissue and cause a reticular fluorescence pattern. With the thyroglobulin-coated BIOCHIP, autoantibodies against thyroglobulin (TG) can be confirmed monospecifically in one and the same test procedure. This BIOCHIP Mosaic can be supplemented as required with further substrates, e.g. rat kidney, to achieve a differentiation of antibodies against thyroid microsomes from mitochondrial antibodies (AMA). For a serological diagnosis of autoimmune polyendocrinopathies, BIOC
Thyroid gland, unfixed and thyroglobulin BIOCHIP: antibodies against thyroid microsomes and thyroglobulin.
Order No. FA 1010-####-3
Formats page 116
Radioimmunoassays (RIA/IRMA): Thyroid Specific Autoantibodies, Antigens and Hormones

Monospecific detection of autoantibodies against thyroglobulin (TG), thyroid peroxidase (TPO) and thyrotropin receptor (TSH-R). Specific detection of the thyroid antigen thyrotropin and the hormones free triiodothyronine (FT3), free thyroxine (FT4), thyrotropin (TSH), calcitonin. Indications: Basedows disease, Hashimoto autoimmune thyroiditis, medullary thyroid carcinoma, thyroidal C-cell hyperplasia, therapy controls in hyper- and hypothyrosis. Serum dilutions: 1:50 for anti-TG and anti-TPO (magnetic), 1:20 for anti-TG and anti-TPO (precipitation), undiluted for all remaining test kits. 5-point to 8-point calibration (quantitative). Analyte Anti-TPO Anti-TG TRAb Thyrotropin Order No. RA 1012-####-# RA 1013-10001-# RA 1015-10001 RD 1013-10001 Analyte FT3 FT4 TSH Calcitonin Order No. RD 1016-10001 RD 1017-10001 RD 1018-10001 RD 1019-10001
RIA for thyroid diagnostics.
Order No. (Anti-TPO) RA 1012-####-#
Formats page 89
Microplate ELISA: Anti-Thyroglobulin, Anti-Thyroid Peroxidase, Anti-TSH receptor

Monospecific determination of antibodies against thyroglobulin (TG), thyroid peroxidase (TPO), and thyrotropin receptor (TSH-R). Indications: Basedows disease, Hashimoto autoimmune thyroiditis. Serum dilution 1 : 200 (exception: anti-TSH-R undiluted); conjugate class antihuman IgG, POD-labelled (anti-TSH-R: avidin-labelled). 3-point calibration (exception: anti-TSH-R, 5-point calibration). The quantification is carried out according to international reference preparations (anti-TG: NIBSC 65/93; anti-TPO: NIBSC 66/387; anti-TSH-R: NIBSC 90/672). Thyroglobulin/TSH-R: native antigen; thyroid peroxidase: recombinant antigen. Antigen Thyroid peroxidase Thyroglobulin TSH receptor Order No. (Anti-TPO) EA 1012-9601 G Formats page 86 Order No. EA 1012-9601 G EA 1013-9601 G EA 1015-9601 G
Incubated ELISA Anti-Thyroid antigens.
TSH receptor (Fast-ELISA) EA 1015-9601-1 G
40
EUROIMMUN
Autoantibodies against Antigens of the Skin

Indirect Immunofluorescence test: Dermatology Mosaic 9

Screening and differentiation test for detection of skin-specific antibodies. Indication: autoimmune bullous dermatoses. Initial dilution 1 : 10; conjugate class anti-human IgG, FITC-labelled. The BIOCHIP Mosaic consists of 6 substrates:primate oesophagus,desmoglein 1expressing cells, desmoglein 3-expressing cells, BP230-expressing cells (whole length), BP230-expressing cells (gC) and BP180 (EUROPLUS). Thus a comprehensive antigen spectrum is available in a single analysis, allowing targeted serological diagnosis. Autoantibodies against intercellular target structures (prickle cell desmosomes) can be reliably detected using tissue sections of oesophagus and tongue, although with this combination it is difficult to distinguish between desmoglein 1 and desmoglein 3. When specific transfected cells are employed in addition, a targeted diagnosis in a single test run is possible. Antibodies against prickle cell desmosomes Antibodies against prickle cell desmosomes react with surface antigens of keratinocytes. Tissue sections of oesophagus and tongue show a granular fluorescence in the intercellular matter in the whole stratum spinosum. When autoantibodies against BP180 or BP230 are present, the epidermal basement membrane in the oesophagus or tongue is visible as a fine linear colouring between the stratum basale and the connective tissue. These antibodies can be differentiated by means of BP180-NC16A-4X coated BIOCHIPS and cells transfected with BP230 (whole length or globular C-terminal domain (gC), respectively. This BIOCHIP Mosaic can be customised with further substrates if required, e.g. tongue (antibodies against prickle cell desmosomes, epidermal basement membrane), bladder (antibodies against plakins), salt split skin (antibodies against epidermal basement membrane). Substrate Desmoglein 1 (transfected / non-transfected cells) Desmoglein 3 (transfected / non-transfected cells) Oesophagus Oesophagus / Tongue Tongue Bladder mucosa Salt split skin Order No. FA 1495-####-50 FA 1496-####-50 FA 1501-#### FA 1501-####-1 FA 1502-#### FA 1507-#### FA 150b-#### Oesophagus: ab against epid. basal membrane (top left). Desmoglein 1 + 3 (middle left, bottom left). BP230 gC (top right). BP230 whole length (middle right). BP180 (NC16A-4X) (bottom right). Order No. FA 1501-####-9 G Formats page 125
Microplate ELISA: Anti-Desmoglein 1, Anti-Desmoglein 3, AntiBP180-NC16A-4X, Anti-BP230-CF

Monospecific detection of antibodies against desmoglein 1, desmoglein 3, BP180 und BP230. Indication: autoimmune bullous dermatoses. Serum dilution 1 : 100; conjugate class anti-human IgG, POD-labelled. 3-point calibration, quantitative. Identical incubation conditions and times: all tests can be combined on one microplate. Recombinant antigens: extracellular domain of desmoglein 1 or 3, tetramer of NC16A domain of BP180 protein, C-terminal segment of BP230 protein. The corresponding human cDNA is produced in E. coli (BP180-NC16A-4X, BP230-CF) or in mammalian cells (desmoglein 1, desmoglein 3). Incubated ELISA Anti-Desmoglein 1 and 3, AntiBP180-NC16A-4X, Anti-BP230-CF Order No. (Anti-Dsg-1) EA 1495-4801 G Formats page 87
41
EUROIMMUN
Autoantibodies against Neuronal Antigens

Indirect Immunofluorescence Test: BIOCHIP Mosaic Cerebellum/ Peripheral Nerves/Intestinal Tissue
Screening test for the detection of antibodies against neuronal antigens. Indications: neurological diseases. Initial dilution 1 : 10; conjugate class anti-human IgAGM, FITC-labelled. Primate cerebellum and primate nerves are the standard substrates for the determination of various neuronal antibodies. The parallel use of primate intestine permits the reliable differentiation from other autoantibodies (e.g. ANA) and makes it possible to distinguish between anti-Ri and anti-Hu. Antibodies against grey matter react intensively with the stratum granulosum and in a weaker form with the stratum moleculare of the cerebellum. Target antigen: glutamic acid decarboxylase (GAD). Clinical significance: stiff person syndrome, diabetes mellitus type I. Antibodies against Yo stain exclusively the cytoplasm of the Purkinje cells in the cerebellum. Clinical significance: paraneoplastic neurological syndromes (PNS), indication of a malignoma. In the case of antibodies against Hu and Ri all neurone nuclei in the grey matter show a granular fluorescence. Hu antibodies react in the intestine with cell nuclei of the plexus myentericus, whereas Ri antibodies do not. Clinical significance: paraneoplastic neurological syndromes (PNS), indication of a malignoma. The white matter of the cerebellum is stained by antibodies against myelin, which present as hyaline cylinders in tissue sections of peripheral nerves. A droplike ring-shaped fluorescence is observed in cross sections of nerves. The fluorescence of the Virchow-Robin space (cerebellum, optic nerve) and the pia is caused by autoantibodies developed in neuromyelitis optica (NMO-IgG). Antibodies against myelin-associated glycoprotein (MAG), on the other hand, show a streaky fluorescence pattern on nerve tissue and a mostly fine-granular ring-shaped fluorescence on cross sections of peripheral nerves. Clinical significance: paraproteinaemic neuropathy. The BIOCHIP Mosaic can be supplemented as required with further substrates, e.g. cerebrum (antibodies against astrocytes), optic nerve, primate liver plus HEp-2 cells (to rule out ANA), Crithidia luciliae (anti-dsDNA), primate stomach (parietal cell antibodies), Borrelia (neuroborreliosis-associated). Aquaporin4(AQP4) transfected HEK cells allow a monospecific antibody determination in suspected cases of neuromyelitis optica (NMO).
Cerebellum: antibodies against GAD and Yo (top), against Hu and Ri (middle). Peripheral nerves: antibodies against myelin and MAG (bottom). Order No. FA 1111-####-1 Formats page 117
amphiphysin CV2.1* PNMA2 (Ma2/Ta) Ri Yo Hu
EUROLINE: Neuronal Antigens Profile 2

Differentiation of antibodies against neuronal antigens. Indication: paraneoplastic neurologic syndromes (PNS). Serum dilution 1 : 100; conjugate class anti-human IgG, AP-labelled. With the EUROLINE Neuronal Antigens Profile 2, six autoantibodies can be determined: antibodies against amphiphysin, CV2.1*, PNMA2 (Ma2/Ta), Ri, Yo and Hu. Test strips can be automatically incubated and evaluated using the systems EUROBlotMaster und EUROLineScan (see page 27). Additionally, EUROIMMUN offers a Westernblot for the detection of antibodies against neuronal antigens: DW 1111-1601 G.
control
*) CV2 partial protein, which only contains the N-terminally localised epitopes of the antigen.
Incubated EUROLINE Neuronal Antigens Profile 2.
Order No. DL 1111-1601-2 G
Formats page 80
42
EUROIMMUN
Autoantibodies against Neuronal Antigens

Indirect Immunofluorecence test: BIOCHIP Mosaic Hippocampus/ Cerebellum/NMDA Receptor

Screening test for detection of antibodies against glutamate receptors (type NMDA, anti-N-methyl-D-aspartate receptor) und neuropil. Indication: NMDAR encephalitis. Initial dilution 1 : 10; conjugate class anti-human IgG, FITC-labelled. Immunohistochemistry with tissue sections of rat hippocampus and rat cerebellum allows identification of antibodies against glutamate receptors (type NMDA) and other antibodies (e.g. VGKC and AMPA receptors). The parallel use of recombinant HEK293 cells enables sensitive and monospecific detection of anti-glutamate receptor (type NMDA) antibodies and their differentiation from other neuropil antibodies in a simple and efficient analysis. Antibodies against glutamate receptors (type NMDA) stain the neuropil of the molecular layer of the hippocampus as well as the granular layer of the cerebellum. They show a flat, smooth to fine-granular fluorescence in the cytoplasm of the transfected HEK293 cells. Clinical significance: Anti-NMDA receptor encephalitis. If the monospecific detection of antibodies against glutamate receptors of type NMDA is negative, neuropil fluorescence can indicate the presence of other antibodies associated with limbic encephalitis (e.g. anti-VGKC antibodies, antiAMPA receptor antibodies).
Antibodies against glutamate receptor (type NMDA): rat hippocampus (top left), rat cerebellum (bottom left), untransfected cells (top right), transfected cells (bottom right).
Order No. FA 111m-####-3 G
Formats page 118
EUROLINE-WB: Anti-Neuronal Antigens

Determination of human autoantibodies against neuronal antigens. Indication: paraneoplastic neurological syndromes (PNS). Serum dilution 1 : 51; conjugate class anti-human IgG, AP-labelled. EUROLINE-WB is a combination of westernblot and line blot techniques. Proteins of a primate cerebellum extract are electrophoretically separated according to molecular mass and transferred onto a nitrocellulose membrane (westernblot). A membrane chip coated with highly purified recombinant Hu, recombinant Ri and recombinant Yo is subsequently applied to the westernblot strip. Test strips can be automatically incubated using the system EUROBlotMaster (Seite 27). Yo Yo
Ri Ri Hu
rec. Yo Control
rec. Ri rec. Hu
Incubated Anti-Neuronal Antigens EUROLINE-WB. Order No. DW 1111-####-2 G Formats page 83
43
EUROIMMUN
Autoantibodies against Islet Cell Antigens

Indirect Immunofluorescence Test: Primate Pancreas
Detection of antibodies against islet cells. Indications: Differentiation between a late manifestation of diabetes type 1 (latent autoimmune diabetes in adulthood, LADA) and diabetes type 2. For a reliable determination of antibodies against islet cells an extended incubation time of 18 hours for the patient serum must be observed. The incubation time may be reduced to 2 hours but this will lead to a decrease in the sensitivity of the antibody detection test. Standardised control with JDF units available (order no. CA 1021-0101-1). With indirect immunofluorescence autoantibodies against pancreas islets (ICA) can be detected in 80% of patients with new-onset diabetes type 1. Two target antigens of ICA have been identified so far: the enzymes glutamic acid decarboxylase (GAD) and tyrosine phosphatase (IA2). This BIOCHIP may be supplemented with further substrates, e. g. primate cerebellum for the detection of antibodies against GAD. The microscopic evaluation can be significantly simplified by using small BIOCHIPs (1 x 1 mm). The BIOCHIPs appear almost completely in the field of view and facilitate finding the islet cells, thus rendering a time-consuming search unnecessary, especially in negative samples.
Primate pancreas: antibodies against islet cells.
Order No. FA 1020-####
Formats page 116
Microplate ELISA: Anti-GAD, Anti-IA2, Anti-GAD/IA2 Pool

Monospecific detection of antibodies against glutamic acid decarboxylase (GAD), tyrosine phosphatase (IA2) or bispecific detection of both antibodies in a single reagent well. Indications: early diagnosis of diabetes mellitus type 1, risk prediction in first grade relatives, prognosis of the clinical progression of diabetes type 1 for prediction of insulin dependence, differential diagnosis in gestational diabetes, differentiation between a late manifestation of diabetes type 1 (latent autoimmune diabetes in adulthood, LADA) and diabetes type 2. Use of undiluted samples. Similar incubation conditions and times. Manual or automated test performance. Multipoint calibration. The quantitation is based on an international reference preparation (NIBSC 97/550). GAD and IA2: human, recombinant antigens. Antigen Order no.
Incubated ELISA anti-GAD.
Glutamic acid decarboxylase (GAD)EA 1022-9601 G Order No. (Anti-GAD) EA 1022-9601 G Formats page 86 Tyrosine phosphatase (IA2) GAD/IA2 Pool EA 1023-9601 G EA 1022-9601-1 G
RIA: Anti-GAD, Anti-IA2, Anti-Insulin

Monospecific detection of antibodies against glutamic acid decarboxylase (GAD), tyrosine phosphatase (IA2) and insulin. Indications: Early diagnosis of diabetes mellitus type 1, risk prediction in first grade relatives, prognosis of the clinical progression of diabetes type 1 for prediction of insulin dependence, differential diagnosis in gestational diabetes, differentiation between a late manifestation of diabetes type 1 (latent autoimmune diabetes in adulthood, LADA) and diabetes type 2. Use of undiluted samples. Similar incubation conditions and times. Manual or automated test performance. Test kit formats for 50 or 100 determinations. GAD and IA2: human, recombinant, 125I-labelled antigens, insulin: human, synthetic, 125I-labelled antigen. Antigen Glutamat-Decarboxylase (GAD) Tyrosin-Phosphatase (IA2) Order No. (Anti-IA2) RA 1023-#### Formats page 89 Insulin Order no. RA 1022-#### RA 1023-#### RA 1024-####
RIA Anti-IA2.
44
EUROIMMUN
Autoantibodies against Parietal Cells (PCA)

Indirect Immunfluorescence Test: Primate Stomach with Urea Pretreatment

Screening test for detection of antibodies against parietal cells. Indications: forms of chronic atrophic gastritis, pernicious anemia, funicular myelosis, various autoimmune endocrinopathies such as Basedows and Addisons diseases. Initial dilution 1 : 10; polyvalent conjugate anti-human IgAGM, FITC-labelled. Primate stomach is the standard substrate for detection of parietal cell antibodies. For titration, stomach tissue from rat or mouse is sufficient. With positive results the parietal cells show a course granular to clumpy fluorescence, and the surrounding areas are usually dark. Parietal cell antibodies (PCA) are often mixed up with antibodies against mitochondria (AMA) in microscopic analysis. The latter give an even fine granular fluorescence of the parietal cell cytoplasm, with the surrounding region showing a (weaker) reaction. For reliable differentiation of both types of antibody, a 30-minute pretreatment of the tissue sections with urea-glycine buffer (order no. ZF 1140-0101, see page 151) is recommended. The cytomplasmic fluorescence of parietal cells resulting from PCA occurs with the same intensity with or without urea pretreatment. The typical pattern of mitochondrial antibodies is almost completely supressed by urea pretreatment, greatly facilitating PCA diagnostics. In some AMA-positive samples it is possible to detect PCA that are obscured by the AMA pattern in conventional tissue sections. The urea-pretreated tissue shows a significantly darker background, enabling specific fluorescence to be more easily and reliable identified. This BIOCHIP can be supplemented with additional substrates, for example, thyroid (thyroid peroxidase, thyroglobulin), pancreas (pancreas islets), adrenal gland (adrenal cortex), ovary (ovary antigens), testis (Leydig cells), and intrinsic factor.
Primate stomach: antibodies against parietal cells. With urea-pretreatment (top) and without urea-pretreatment (bottom).
Primate stomach: antibodies against mitochondria. With urea-pretreatment (top) and without urea-pretreatment (bottom).
Order No. FA 1360-#### G
Formats page 122
Microplate ELISA: Anti-Parietal Cells

Monospecific detection of antibodies against parietal cells (PCA). Indications: forms of chronic atrophic gastritis, pernicious anemia, funicular myelosis, various autoimmune endocrinopathies such as Basedows and Addisons diseases Serum dilution 1 : 200; conjugate class anti-human IgG, POD-labelled. 3-point calibration, quantitative. Native antigen: H+/K+-ATPase, purified by affinity chromatography.
Incubated ELISA Anti-Parietal Cells.
Formats page 86
45
EUROIMMUN
Autoantibodies against Granulocyte Cytoplasm (cANCA/pANCA)

Indirect Immunofluorescence Test: EUROPLUS Granulocyte Mosaic 23
Screening test for the detection of antibodies against granulocyte cytoplasm (ANCA). Indications: Wegeners granulomatosis, various forms of glomerulonephritis, primary sclerosing cholangitis, ulcerative colitis, Crohns disease. Initial dilution: serum 1 : 10; conjugate anti-human IgG, FITC-labelled. Using ethanol-fixed granulocytes, antibodies against granulocyte cytoplasm can be detected. In this case, two fluorescence patterns have to be differentiated: a granular fluorescence which is distributed evenly over the entire cytoplasm, leaving the cell nuclei free (cytoplasmic type, cANCA) or a smooth fluorescence wrapped ribbon-like around the cell nuclei (perinuclear type, pANCA). Antibodies against all relevant granulocyte antigens as well as against further, as yet unknown antigens are detected simultaneously: Pattern cANCA pANCA pANCA pANCA pANCA pANCA Target antigen Proteinase 3 Myeloperoxidase Elastase Cathepsin G Lysozyme Lactoferrin Associated diseases Wegeners granulomatosis Microscopic arteritis, Churg-Strauss syndrome, polyarteritis nodosa Ulcerative colitis, Crohns disease, primary sclerosing cholangitis, systemic lupus erythematosus Ulcerative colitis, primary sclerosing cholangitis, Crohns disease Ulcerative colitis, primary sclerosing cholangitis, Crohns disease Ulcerative colitis, primary sclerosing cholangitis, Crohns disease, systemic lupus erythematosus, rheumatoid arthritis Primary sclerosing cholangitis, ulcerative colitis, Crohns disease Ulcerative colitis, Crohns disease
cANCA BPI or pANCA pANCA unknown
EUROPLUS Granulocyte Mosaic 23 (granuloc. (EOH), granuloc. (HCHO), PR3, MPO, HEp-2, primate liver): pattern pANCA, formalin-resistant. Order No. FA 1201-####-23 Formats page 120
The primate liver enables one to differentiate between pANCA and anti-nuclear antibodies (ANA) which can easily be confused when using ethanol-fixed granulocytes: In the case of a positive ANA result all nuclei of the hepatocytes fluoresce, whereas in the case of pANCA (as well as cANCA) only the granulocytes in the sinusoids of primate liver fluoresce. The EUROPLUS substrates PR3 and MPO as monospecific tests can confirm results from conventional granulocyte screening tests. Recombinant GBM EUROPLUS substrate also provides additional reliability for diagnosis. When fluorescence patters are unclear (e.g. unspecific fluorescence caused by other cytoplasmic antibodies) these substrates facilitate evaluation.
Microplate ELISA: ANCA Profile

Differentiation of antibodies against granulocyte cytoplasm (ANCA). Indications: Wegeners granulomatosis, various forms of glomerulonephritis, primary sclerosing cholangitis, ulcerative colitis, Crohns disease. Serum dilution 1 : 100; conjugate class anti-human IgG, POD-labelled. 6 relevant anti-granulocyte antibodies can be detected simultaneously and monospecifically: autoantibodies against proteinase 3, lactoferrin, myeloperoxidase, elastase, cathepsin G, BPI. 1-point calibration, semi-quantitative. Calibrator pool and serum sample on the same microplate strip. Native antigens, purified by affinity chromatography. Available individual ELISA (3-point calibration, quantitative): Antigen Proteinase 3 (Capture ELISA) Order No. EA 1201-9601-1 G
Incubated ELISA ANCA Profile (antigens: proteinase 3, lactoferrin, myeloperoxidase, elastase, cathepsin G, BPI.
Order No. EA 1200-1208-1 G
Formats page 86
Proteinase 3 (PR3-hn-hr: native/recombinant) EA 1201-9601-2 G Myeloperoxidase EA 1211-9601 G
46
EUROIMMUN
Cost-effective Strategy for the Detection of Autoantibodies against Granulocyte Cytoplasm (ANCA)
Screening test indirect immunofluorescence: BIOCHIP sextet granulocytes (EOH), granulocytes (HCHO), EUROPLUSTM microdots PR3, EUROPLUSTM microdots MPO, human epithelial cells (HEp-2), and primate liver
Perinuclear fluorescence of granulocytes (A, F) A Granulocytes (EOH-fixed) F Primate liver Fluorescence of all cell nuclei (A, E, F) Fluorescence of all cell nuclei (A, E, F), granuloc. accent. (F) Cytoplasmic fluorescence of granulocytes (A, B, F)
B Granulocytes (HCHO-fixed)
E HEp-2 cells
C EUROPLUS PR3 microdots

TM
D EUROPLUS MPO microdots

TM
pANCA MPA 42-70 % CU CSS 18-60 % MC SLE 9-25 % PSC RA 3-25 % 67 % 7% 87 %
only ANA
ANA and pANCA WG MPA CSS PAN
cANCA 80-90 % 10-15 % 10-20 % <9%
At B cytoplasmic fluorescence
At B no cytoplasmic fluorescence
Cytoplasmic fluorescence of granulocytes (B)
Nuclei of granuloc. brighter than nuclei of hepatocytes (F), B neg.
ANCA reaction at A, ANA reaction at E & F
pANCA, formalin-resistant
pANCA, formalin-sensitive
pANCA, formalin-res. & ANA MPA 42-70 %
pANCA, formalin-sens. & ANA
pANCA, formalin-sens.? & ANA
ELISA: anti-MPO AAb against MPO
ELISA: ANCA Profile AAb against PR3, MPO, elastase, cathepsin G, BPI, lactoferrin
Qualified ANA diagnostics: screening test using HEp-2 cells/primate liver (E, F), differentiation using ELISA, EUROASSAY, EUROLINE, Westernblot
ELISA: anti-PR3-hn-hr / ANCA Profile (BPI) AAb against PR3-hn-hr AAb against BPI
MPA
53 %
WG 5 % (BPI) MPA 53 % (MPO) MPA 3 % (LF) RA, vasculitides 45 % (LF) SLE 6 % (EL)
AAb against dsDNA, ssDNA, nucleosomes, histones, nuclear membrane, nRNP/Sm, Sm, SS-A, Ro-52, SS-B, Ku, cyclin I (PCNA), mitosin (CENP-F, cyclin II), nuclear dots, centromeres (CENP B), spindle fibres, midbody, centrioles, Scl-70, PM-Scl, fibrillarin, RNA polymerase I, NOR, ribosomal P-proteins, Jo-1, PL-7, PL-12, Mi-2, mitochondria (AMA), lysosomes, Golgi apparatus, vimentin, tropomyosin, actin.
WG
80-90 %
WG
5%
See EUROIMMUN poster: "Strategy for Determination of Autoantibodies against Cell Nuclei (ANA) and Cytoplasm Components"
The highest diagnostic sensitivity in the determination of autoantibodies against neutrophil granulocytes (ANCA) is achieved by using indirect immunofluorescence and assays with defined target antigens (particularly PR3 and MPO) simultaneously at the start. However, under the pressure of cost optimisation, an immunofluorescence test may be performed on its own and then followed up by specific ELISA tests only if the result is positive. Ethanol-fixed human granulocytes are the standard substrate for indirect immunofluorescence. With this substrate two relevant fluorescence patterns can be differentiated: the cytoplasmic type (cANCA) associated with Wegeners granulomatosis and the perinuclear type (pANCA), which indicates a range of various diseases. The differentiation of pANCA from antibodies against cell nuclei (ANA) is often difficult. Therefore, HEp-2 cells (possibly with sedimented granulocytes) or primate liver are used as an additional substrate. If ANA and pANCA occur together, the granulocytes show a much brighter fluorescence than the cell nuclei. Thanks to EUROIMMUN BIOCHIPs it is not necessary to incubate human epithelial cells on a second slide in parallel for the exclusion of cell nucleus antibodies, since all substrates are present in one and the same test field. A fourth BIOCHIP with formalin-fixed human granulocytes detects a large proportion of the diagnostically relevant antibodies against myeloperoxidase, whereas other pANCA (which are particularly important in gastroenterology) and almost all antibodies against cell nuclei (whose differentiation is a separate chapter in autoantibody diagnostics) are generally completely suppressed. The EUROPLUSTM substrates PR3 and MPO help to confirm diagnosis and allow a quick and reliable interpretation of results even in problematic cases.
ANA: anti-nuclear antibodies BPI: bactericidal permeability increasing protein cANCA: anti-neutrophil cytoplasmic antibodies, cytoplasmic type CD: Crohns disease CSS: Churg-Strauss syndrome EL: elastase EOH: ethanol HCHO: formalin HEp-2: human epithelial cells IFT: immunofluorescence test LF: lactoferrin MPA: microscopic polyangiitis MPO: myeloperoxidase PAN: polyarteritis nodosa pANCA: anti-neutrophil cytoplasmic antibodies, perinuclear type PR3: proteinase 3 PSC: primary sclerosing cholangitis RA: rheumatoid arthritis SLE: systemic lupus erythematosus UC: ulcerative colitis WG: Wegeners granulomatosis
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EUROIMMUN
Antibodies against Endomysium and Gliadin

Indirect Immunofluorescence Test: EUROPLUS Primate Liver and Gliadin (GAF-3X) BIOCHIPs
Detection of antibodies against endomysium and gliadin. Indications: gluten-sensitive enteropathy (celiac disease, non-tropical sprue), Duhrings herpetiform dermatitis. Initial dilution 1 : 10; conjugate class anti-human IgA or IgG, FITC-labelled. Autoantibodies against endomysium react with many types of tissue, e.g. primate oesophagus. The most suitable substrate, however, is primate liver: in the case of a positive sample, filamentous linings of the intralobular sinusoids react. With the gliadin (GAF-3X)-coated BIOCHIP, antibodies against gliadin can be analyzed in one and the same test procedure. Both anti-endomysium antibodies and antibodies against gliadin (class IgA) are reliable serological markers for an active gluten-sensitive enteropathy. Therefore, their determination can in many cases replace endoscopy and biopsy.
Primate liver and gliadin (GAF-3X) BIOCHIP: antibodies against endomysium and gliadin.
Order No. FA 1914-####-1 A or G
Formats page 132
Microplate ELISA: Anti-Gliadin (GAF-3X)

Monospecific detection of antibodies against gliadin. Indications: gluten-sensitive enteropathy (celiac disease, non-tropical sprue), Duhrings herpetiform dermatitis. Serum dilution 1 : 200; conjugate class anti-human IgA or IgG, POD-labelled. 3-point calibration. Identical incubation conditions and times: the investigation of IgA and IgG antibodies can be combined without difficulty on one and the same microplate. Antigen: Gliadin-analogue fusion peptide (GAF-3X). The quantitative determination of antibodies against gliadin is very suitable for monitoring the progress of the disease, compliance with a gluten-free diet, or a gluten tolerance test.
Incubated ELISA Anti-Gliadin (GAF-3X).
Order No. EV 3011-9601 A or G
Formats page 95
Microplate ELISA: Anti-Tissue Transglutaminase (Endomysium)

Monospecific detection of antibodies against tissue transglutaminase. Indications: gluten-sensitive enteropathy (celiac disease, non-tropical sprue), Duhrings herpetiform dermatitis. Serum dilution 1 : 200; conjugate class anti-human IgA or IgG, POD-labelled. 3-point calibration, quantitative. Antigen: recombinant, expression with the baculovirus vector in insect cells.
Incubated ELISA Anti-Tissue Transglutaminase (Endomysium).
Order No. EA 1910-9601 A or G
Formats page 88
48
EUROIMMUN
EUROIMMUN PRODUCTS FOR INFECTIOUS SEROLOGY
49
EUROIMMUN Products for Infectious Serology
EUROIMMUN
Antibodies against Borrelia

Indirect Immunofluorescence Test: EUROPLUS Anti-Borrelia afzelii, Borrelia burgdorferi, VlsE and OspC Antigen

Antibodies against Borrelia afzelii, VlsE (top), Borrelia burgdorferi and OspC (bottom). Order No. FI 2136-####-1 G or M Formats page 136
Sensitive screening test for the detection of anti-Borrelia antibodies. Indications: erythema chronicum migrans, lymphadenosis cutis benigna, lymphocytic meningoradiculitis, carditis, arthritis, acrodermatitis chronica atrophicans, neuroborreliosis. Initial dilution 1 : 10 (IgM), 1 : 100 (IgG). If antibodies against Borrelia afzelii or Borrelia burgdorferi are present, a distinct fluorescence of the bacteria in the smear is obtained. With the VlsE or OspC coated BIOCHIPs antibodies against the highly specific and highly sensitive marker antigens VlsE (IgG) or OspC (IgM) can be determined monospecifically in one and the same test procedure. If these antigens fluoresce the antibody result is positive even if the bacteria smears show a negative reaction. Thus, the BIOCHIP Mosaic helps to increase specificity and sensitivity in Borrelia diagnostics. The BIOCHIP can be supplemented as required using further substrates, e.g. Borrelia burgdorferi sensu stricto (strains CH or USA) and TBE virus infected cells.
Microplate ELISA: Anti-Borrelia plus VlsE

Sensitive screening test for the detection of anti-Borrelia antibodies. Indications: erythema chronicum migrans, lymphadenosis cutis benigna, lymphocytic meningoradiculitis, carditis, arthritis, acrodermatitis chronica atrophicans, neuroborreliosis. Serum dilution 1 : 100; conjugate class anti-IgG, anti-IgM or anti-IgAGM (VlsE: IgG only), POD-labelled. 3-point calibration (IgG and IgM). Identical incubation conditions and times: all tests can be combined without difficulty on one and the same microplate. Antigens: extract of Borrelia burgdorferi sensu stricto, Borrelia garinii and Borrelia afzelii (whole antigen) / recombinant VlsE from Borrelia burgdorferi sensu stricto. VlsE (variable major protein-like sequence, expressed) is a newly characterized surface protein of Borrelia which is expressed exclusively in vivo and which contains conserved and highly immunogenic epitopes. IgM test kit (Anti-Borrelia) includes IgG/RF absorbent in sample buffer for IgG absorption in preparation for the determination of specific IgM class antibodies.
Incubated ELISA Anti-Borrelia plus VlsE.
Order No. EI 2132-9601 G or M
Formats page 91
Microplate ELISA: Anti-Borrelia plus VlsE, Antibody Determination in Serum and Cerebrospinal Fluid for Detection of Intrathecal Synthesis of Specific Antibodies against Borrelia
Antibody determination in serum and cerebrospinal fluid (CSF). Indication: Neuroborreliosis. CSF dilution 1 : 2, serum dilution 1 : 404; conjugate class anti-IgG, POD-labelled. 4-point calibration, quantitative. Microplate ELISA for the detection of Borrelia antibodies of class IgM in serum and CSF are also available.
Table-based evaluation of the CSQrel.
Order No. EI 2132-9601 L G
Formats page 91
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EUROIMMUN
Antibodies against Borrelia

Anti-Borrelia EUROLINE-RN-AT
Specific confirmatory test for the detection of antibodies against Borrelia. Indications: erythema chronicum migrans, lymphadenosis cutis benigna, lymphocytic meningoradiculitis, carditis, arthritis, acrodermatitis chronica atrophicans, neuroborreliosis. The Anti-Borrelia EUROLINE is coated with both native and recombinant proteins and provides a unique mixture of Borrelia specific antigens: The classical antigens OspC, p83 and p39, which show the highest specificity in their native form, were accurately cut from a Westernblot and applied onto the membrane of the line blot. Five new, recombinant designer antigens (p18, p19, p20, p21, p58) having a very high specificity (IgG) were identified using bioinformatic analysis of the Borrelia genome. For the first time, lipids which have been proven to be immunoreactive and were extracted from the Borrelia membranes, three native OspC antigens (IgM) from B. afzelii, B. burgdorferi and B. garinii and three different VlsE antigens (IgG) from B. afzelii, B. burgdorferi and B. garinii are available on the line blot. The serological hit rate is increased by 10% by using three OspC variants in the IgM test. Serum dilution 1 : 50; conjugate class anti-IgG or anti-IgM, AP-labelled.
VlsE B. burgd. p41 B. afzelii p39 B. afzelii OspC B. afzelii OspC B. burgd. OspC B. garinii VlsE B. afzelii VlsE B. burgdorferi VlsE B. garinii Lipid B. afzelii Lipid B. burgdorferi p83 B. afzelii p41 B. garinii p39 B. garinii OspC B. garinii p58 p21 p20 p19 p18 IgM Control band IgG Control band
Incubated Anti-Borrelia EUROLINE-RN-AT . Order No. DN 2131-3201 G or M Formats page 81
EUROLINE-WB: Anti-Borrelia (Whole Antigen plus VlsE)

Specific confirmatory test for the detection of antibodies against Borrelia. EUROLINE-WB is a combination of westernblot and line blot techniques. An SDS extract of a Borrelia afzelii strain is electrophoretically separated according to molecular mass and transferred onto a nitrocellulose membrane. A membrane chip coated with highly purified recombinant VlsE-Antigen is then added to the westernblot strips. By additionally determining antibodies against VlsE the serological hit rate can be increased by 20% compared to whole extract Westernblots and by 30% compared to recombinant antigen Westernblots. Of all recombinant antigens, VlsE possesses the highest sensitivity for the detection of a Borrelia infection. Over 85% of IgG-positive sera could be identified at a glance by assessing the VlsE band. VlsE allows detection of antibodies against all Borrelia species, and the risk of a false negative reaction due to species differences is ten times lower. Serum dilution 1 : 50; conjugate class anti-IgG or anti-IgM, AP-labelled.
VlsE p 83
p 41, Flag. p 39, Bmp A p 31, Osp A p 30 p 25, Osp C p 21 p 19 p 17
Control band
Incubated EUROLINE-WB Anti-Borrelia. Order No. DY 2131-1601-1 G or M Formats page 84
Automated Evaluation of Incubated Membrane Strips with the System EUROLineScan
The program EUROLineScan from EUROIMMUN has been developed to enable quantitative evaluation of membrane based test systems, facilitate management of data, and provide detailed documentation of results tasks which have until now required considerable time. First, the incubated test strips are scanned using a flatbed scanner or camera system. EUROLineScan recognizes the position of the strips, even if they have been placed inexactly, identifies the bands, and measures their intensity. The automated evaluation can be monitored, and it is possible to supplement the data manually. The results are then saved together with the image data. It is no longer necessary to archive (potentially infectious) incubated test strips. A separate results sheet can be produced for each patient.
51
EUROIMMUN
Antibodies against Epstein-Barr Virus (EBV)
Patient 1: EBV-CA (IgG)
EBV-CA (IgG): urea-treated
EBV-CA (IgM)
EBV-EA (IgG)
EBNA (IgG)
Patient 2: EBV-CA (IgG)
EBV-CA (IgG): urea-treated
EBV-CA (IgM)
EBV-EA (IgG)
EBNA (IgG)
Indirect Immunofluorescence Test: BIOCHIP Sequence EBV

Gold standard for the determination of antibodies against the EBV-CA antigens (Epstein-Barr virus capsid antigen), EBV-EA (EpsteinBarr early antigen) and EBNA (Epstein-Barr nuclear antigen). Indication: infectious mononucleosis, Burkitt's lymphoma, nasopharyngeal carcinoma (NPC). IgG antibodies against EBV-CA indicate an EBV infection. An at least twofold increase in titer and the absence of antibodies against EBNA at the same time is characteristic for the early phase of the infection. IgM antibodies against EBV-CA and antibodies against EBV-EA can also be found in acute infections, but they do not necessarily always occur. The presence of antibodies against EBNA generally indicates the late phase of an EBNA infection. In cases of an acute EBV infection which cannot be reliably discriminated from a relapse or reinfection, the determination of the antibody avidity using a modified immunofluorescence test as an additional parameter is useful. This requires an additional incubation with urea solution (ZF 1130-0801). The determination of low-avidity antibodies against EBV-CA indicates an acute infection. For the monospecific confirmation of EBV-CA antibodies in the same test procedure the BIOCHIP containing ECV-CA is supplemented with the antigen substrates gp125 antigen (native) and p19 antigen (recombinant) (EUROPLUS FI 2791-####-20 G or M). For highly differentiated diagnostics the BIOCHIP Sequence EBV (FI 2799-####-1 X) can be supplemented by using the antigens gp125 and p19 (EUROPLUS FI 2799-####-21 X). Due to the high prevalence of anti-EBV-CA IgA in NPC patients, the parameter is well suited for screening. Confirmation of the result by determination of IgA antibodies against EBV-EA is recommended. Further anti-EBV test kits for indirect immunofluorescence: Substrate Order No. Order No. FI 2799-####-21 X Formats page 148 EBV-CA EBNA FI 2791-#### A, G or M FI 2793-#### G Substrate Order No.
EBV-EA FI 2795-#### A or G EBV-CA & EBV-EA FI 2791-####-2 A or G
Microplate ELISA: Anti-EBV-CA, Anti-EBNA-1, Anti-EBV-EA

Specific confirmatory test for antibodies against EBV-CA, EBNA-1 or EBV-EA. Indications: infectious mononucleosis, Burkitts lymphoma, nasopharyngeal carcinoma. Serum dilution 1 : 100; conjugate class anti-IgA, -IgG or IgM, POD-labelled. 1-point calibration, semi-quantitative (IgA, IgM) or 3-point calibration, quantitative (IgG). Identical incubation conditions and times: all tests can be combined without difficulty on one and the same microplate. EBV-CA: native antigen, purified by affinity chromatography; EBNA-1 and EBV-EA: recombinant antigen. IgM test kit includes IgG/RF absorbent in sample buffer for IgG absorption in preparation for the determination of specific IgM class antibodies. Available individual ELISA: Antigen EBV-CA Order No. (anti-EBNA-1) EI 2793-9601 G Formats page 95 EBNA-1 EBV-EA Order No. EI 2791-9601 A, G or M EI 2793-9601 G EI 2795-9601 A, G or M
Incubated ELISA Anti-EBNA-1.
52
previous infection
fresh infection
EUROIMMUN
Antibodies against Epstein-Barr Virus (EBV)

Determination of Low-Avidity Antibodies against EBV-CA
An alternative principle for the serological diagnosis of fresh infections with EBV has been established by investigating the antibody avidity. The first reaction of the immune system following an infection is the formation of low-avidity antibodies. As the infection proceeds, increasingly antigen-adapted IgG is formed, and avidity grows. As long as high-avidity IgG is not yet detected in the serum, it can be assumed that the infection is still in an early stage. To identify low-avidity antibodies against EBV-CA in a patients serum, two microplate ELISA or immunofluorescence tests are performed in parallel: one test is carried out in the conventional way, the other one includes urea treatment between incubations with patients serum and peroxidase-labelled anti-human IgG, resulting in the detachment of low-avidity antibodies from the antigens. Low-avidity antibodies against EBV-CA are present if the ELISA extinction is significantly reduced by urea treatment. For an objective interpretation, the relative avidity index (RAI) can be calculated out of the measured values with and without urea incubation. With the immunofluorescence test the presence of low-avidity antibodies has been proved if the test using urea treatment gives a far weaker fluorescence than the two-step test.
10
Number of Patients
RAI =
Ewith urea Ewithout urea Primary Infection
Previous Infection
25
50
75
100
Relative Avidity Index (RAI) in %
Avidity of antibodies against EBV-CA (IgG)
Order No. (IIFT) FI 2791-#### X Order No. (ELISA) EI 2791-####-1 G
Formats page 95 and 147
EUROLINE: EBV Profile 2

Differentiation of antibodies against Epstein-Barr virus antigens. Indications: infectious mononucleosis, Burkitts lymphoma, nasopharyngeal carcinoma. Serum dilution 1 : 100; conjugate class anti-human IgG or IgM, AP-labelled. With the EUROLINE EBV Profile 2, five different antibodies can be determined: antibodies against VCA gp125, VCA p19, EBNA-1, p22, EA-D. Recombinant antigens (exception: VCA gp125, native, purified by affinity chromatography). EBNA-1 (IgG) negative und VCA (IgG and IgM) positive: acute (fresh) infection. EBNA-1 (IgG) and VCA (IgG) positive and VCA (IgM) negative: late phase of infection. EBNA-1 (IgG) negative, but band p22 (IgG) and VCA (IgG) positive: late phase of infection with loss of anti-EBNA-1. Test strips can be automatically incubated and evaluated using the systems EUROBlotMaster und EUROLineScan (see page 27).
VCA gp125 VCA p19 EBNA-1 p22 EA-D
Control
Incubated EUROLINE EBV Profile 2.
Order No. DN 2790-1601-2 G or M
Formats page 81
Westernblot: Anti-EBV
Specific confirmatory test for the detection of antibodies against Epstein-Barr virus antigens, differentiation of antibodies against Epstein-Barr virus antigens. Indications: infectious mononucleosis, Burkitts lymphoma, nasopharyngeal carcinoma. Serum dilution 1 : 50; conjugate class anti-IgG or anti-IgM, AP-labelled. Antigens: whole antigen, SDS extract. Bands from all specific antigens are included and clearly separated. EBNA-1 (IgG) negative und VCA (IgG and IgM) positive: acute (fresh) infection. EBNA-1 (IgG) and VCA (IgG) positive and VCA (IgM) negative: late phase of infection. EBNA-1 (IgG) negative, but band p22 (IgG) and VCA (IgG) positive: late phase of infection with loss of anti-EBNA-1. Test strips can be automatically incubated and evaluated using the systems EUROBlotMaster und EUROLineScan (see page 27). VCA 125 EA R EBNA-1 VCA 65 EA D VCA 40/41/42 VCA 33 VCA 22
Control band Incubated Westernblot Anti-EBV. Order No. DY 2790-1501 G or M Formats page 85
53
EUROIMMUN
Antibodies against Helicobacter pylori

Indirect Immunofluorescence Test: Anti-Helicobacter pylori

Sensitive screening test for the detection of antibodies against Helicobacter pylori. Indications: gastritis, ulcus ventriculi et duodeni. Late consequences: MALT lymphomas and adenocarcinomas. Initial dilution 1 : 10 (IgM), 1 : 100 (IgG), 1 : 32 (IgA). If antibodies against Helicobacter pylori are present, a distinct fluorescence of the bacteria in the smear is obtained. A positive IgA result correlates well with the activity of a gastritis. An increased IgG antibody titer is considered to be a marker for chronic infections. A significant drop in the IgG antibody titer about 6 months after therapy is a sign of success. The BIOCHIP can be supplemented as required with further substrates, e.g. other infectious agents or tissue sections of primate stomach.
Antibodies against Helicobacter pylori.
Order No. FI 2080-#### A or G
Formats page 134
Microplate ELISA: Anti-Helicobacter pylori

Sensitive screening test for the detection of antibodies against Helicobacter pylori. Indications: gastritis, ulcus ventriculi et duodeni. Late consequences: MALT lymphomas and adenocarcinomas. Serum dilution 1 : 100; conjugate class anti-IgA or anti-IgG, POD-labelled. Antibodies against Helicobacter pylori antigens can be determined quantitatively in RU/ml. 1-point calibration, semi-quantitative (IgA) or 3-point calibration, quantitative (IgG). Identical incubation conditions and times: both tests can be combined without difficulty on one and the same microplate. Native antigens.
Incubated ELISA Anti-Helicobacter pylori.
Order No. EI 2080-9601 A or G
Formats page 91
p 120, CagA p 95, VacA p 66, UreB
EUROLINE-WB: Anti-Helicobacter pylori

Specific confirmatory test for the detection of antibodies against Helicobacter pylori. Indications: gastritis, ulcus ventriculi et duodeni. Late consequences: MALT lymphomas and adenocarcinomas. Serum dilution 1 : 50; conjugate class anti-IgA or anti-IgG, AP-labelled. EUROLINE-WB is a combination of westernblot and line blot techniques. An SDS extract of a Helicobacter pylori strain is electrophoretically separated according to molecular mass and transferred onto a nitrocellulose membrane (westernblot). Two membrane chips coated with highly purified recombinant CagA and VacA are subsequently applied to the westernblot strips. Bands from all specific antigens are included and clearly separated. Test strips can be automatically incubated and evaluated using the systems EUROBlotMaster und EUROLineScan (see page 27).
p p p p
33 30 29, UreA 26
p 19, OMP p 17 Alignment bar Control band Incubated EUROLINE-WB Anti-Helicobacter pylori. Order No. DY 2080-1601-1 A or G Formats page 84
54
EUROIMMUN
Antibodies against Herpes Simplex Virus (HSV)

Indirect Immunofluorescence Test: BIOCHIP Mosaic HSV-1/HSV-2
Sensitive screening test for the detection of antibodies against herpes simplex viruses. Indication: herpes simplex. Initial dilution 1 : 10 (IgM), 1 : 100 (IgG). If antibodies against herpes simplex-2 virus are present in the sample, a typical fluorescence of the infected cells is obtained mainly in the outspread cytoplasm, less in the cell nuclei. As HSV-1 and HSV-2 are morphologically and immunologically closely related, cross-reactions can occur. Differentiation may be attempted by testing a serum against both antigen substrates and comparing the titers. The BIOCHIP Mosaic can be supplemented as required with further substrates, e.g. other infectious agents. Anti-HSV individual tests for indirect immunofluorescence: Substrate HSV-1 HSV-2 HSV-1 and -2 Order No. FI 2531-#### G or M FI 2532-#### G or M FI 2531-####-1 G or M Order No. FI 2531-####-1 G or M Formats page 141
Antibodies against HSV-1 and HSV-2.
Microplate ELISA: Anti-HSV-1, Anti-HSV-2

Specific confirmatory tests for antibodies against HSV-1 or HSV-2. Indication: herpes simplex. Serum dilution 1 : 100; conjugate class anti-IgG or anti-IgM, POD-labelled. 1-point calibration, semiquantitative (IgM) or 3-point calibration, quantitative (IgG). Identical incubation conditions and times: all tests can be combined without difficulty on one and the same microplate. Antigens: type-specific glycoproteins C1 or G2, purified by affinity chromatography. Cross-reactions do not occur. IgM test kit includes IgG/RF absorbent in sample buffer for IgG absorption in preparation for the determination of specific IgM class antibodies. Available individual ELISA: Antigen HSV-1 HSV-2 HSV-1/2-Pool Order No. EI 2531-9601-2 G or M EI 2532-9601-2 G or M EI 2531-9601-1 G or M Order No. (anti-HSV-1) EI 2531-9601-2 G or M Formats page 93 Incubated ELISA Anti-HSV-1.
EUROLINE-WB: Anti-HSV
Specific confirmatory test for the differentiation of antibodies against HSV-1 and HSV-2. Indication: herpes simplex. Serum dilution 1 : 50; conjugate class anti-IgG or anti-IgM, AP-labelled. EUROLINE-WB is a combination of westernblot and line blot techniques. Proteins from an SDS extract of HSV-1 are electrophoretically separated according to molecular mass and transferred onto a nitrocellulose membrane. A membrane chip coated with HSV-2 type-specific glycoprotein G2 (gG 2), purified by affinity chromatography, is then added to the westernblot strips. Bands from all specific antigens are included and clearly separated. The gG 2 band allows the simple differentiation between HSV-1 and HSV-2 infections. Test strips can be automatically incubated and evaluated using the systems EUROBlotMaster und EUROLineScan (see page 27). Incubated EUROLINE-WB Anti-HSV. Order No. DY 2531-1601-1 G or M Formats page 85
gC 1 gG 1
gG 2 Alignment bar Control band
55
EUROIMMUN
Antibodies against Chlamydia

Indirect Immunofluorescence test: Anti-Chlamydia MIF (MicroImmunofluorescence test)

Serological gold standard for the determination of antibodies against Chlamydia. Indication: trachoma, urogenital infections, lymphogranuloma venereum, laryngitis, sinusitis, bronchitis, pneumonia, psittacosis. Initial dilution 1 : 10 (IgA), 1 : 100 (IgG), 1 : 10 (IgM). The micro-immunofluorescence test uses purified elementary bodies of the species C. trachomatis, C. pneumoniae and C. psittaci as the antigen. The mutual lipopolysaccharide (LPS) antigen is inactivated to minimise cross reactivity. The evaluation of the MIF could be significantly facilitated compared to conventional test systems by using a cell-based substrate. The fourth BIOCHIP with non-infected cells allows a reliable differentiation between unspecific and specific fluorescence.
Anti-Chlamydia MIF.
Order No. FI 2191-####-3 A, G or M
Formats page 138
Microplate ELISA: Anti-Chlamydia trachomatis

Monospecific detection of antibodies against Chlamydia trachomatis. Indication: trachoma, conjunctivitis, urogenital infections, pneumonia in infants, lymphogranuloma venereum. Serum dilution 1 : 100, conjugate class anti-human IgA, IgG or IgM, PODlabelled. 1-point calibration, semiquantitative (IgA and IgM) or 3-point calibration, quantitative (IgG). Antigen: native MOMP antigen (major outer membrane protein). MOMP is a transmembrane protein and represents the main part of the outer membrane of the elementary bodies. Protein purification starts with BGM cells infected with Chlamydia trachomatis of the serotype K. IgM test kit includes IgG/RF absorbent in sample buffer for IgG absorption in preparation for the determination of specific IgM class antibodies.
Incubated ELISA Anti-Chlamydia trachomatis.
Order No. EI 2191-9601 A, G or M
Formats page 92
Microplate ELISA: Anti-Chlamydia pneumoniae

Monospecific detection of antibodies against Chlamydia pneumoniae. Indication: laryngitis, sinusitis, bronchitis, pneumonia. Serum dilution 1: 100, conjugate class anti-human IgA, IgG or IgM, POD-labelled. 1-point calibration, semiquantitative (IgA and IgM) or 3-point calibration, quantitative (IgG). Antigen: cell lysate from HL cells, strain CDC/CWL-029. IgM test kit includes IgG/RF absorbent in sample buffer for IgG absorption in preparation for the determination of specific IgM class antibodies.
Incubated ELISA Anti-Chlamydia pneumoniae.
Order No. EI 2192-9601 A, G or M
Formats page 92
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EUROIMMUN
Antibodies against Emerging Viruses

Indirect Immunofluorescence Test
Over the past years it has been observed that a number of new viruses ("emerging viruses") and other pathogens has spread worldwide, introducing since then unknown diseases into previously unaffected regions. Antibodies against SARS CoV, TBE virus, West Nile virus.
EUROIMMUN offers a broad spectrum of indirect immunofluorescence tests for the detection of specific antibodies against:
Pathogen Disease, syndromes Severe acute respiratory syndrome (SARS) Tick-borne encephalitis West Nile fever, encephalitis Japanese encephalitis Yellow fever, hepatitis, haemorrhagic fever, arthritis Dengue fever, haemorrhagic fever HFRS (haemorrhagic fever with renal syndrome); HCPS (Hantaviral cardiopulmonary syndrome) Pappataci fever, meningitis, encephalitis Rift valley fever, haemorrhagic fever, hepatitis see page 142 143 143 143, 150 143, 144 143, 144, 150 147
Corona virus
SARS corona virus (SARS-CoV) TBE virus (TBEV), West Nile virus (WNV),
Flaviviruses
Japanese encephalitis virus (JEV) Yellow fever virus (YFV) Dengue virus (DENV, types 1-4) Hantavirus (types Hantaan, Puumala, Seoul, Saaremaa, Dobrava, Sin Nombre and Andes) Sandfly fever virus (types Sicilian, Naples, Toscana and Cyprus) Rift valley fever virus (RVFV)
Antibodies against Japanese encephalitis virus, Yellow fever virus, Dengue virus.
146 149 149 150 139 140, 150 140, 150
Bunyaviruses
Antibodies against Sandfly fever virus, Hantavirus, Crimean-Congo fever virus.
Crimean-Congo fever virus (CCHFVCrimean-Congo haemorrhagic fever GPC and -N) Togaviruses Kinetoplastida Haemosporina Plasmodium vivax Malaria tertiana Chikungunya virus (CHIKV) Leishmania donovani Plasmodium falciparum Chikungunya fever, arthritis Visceral leishmaniasis Malaria tropica
Many of these substrates are available as single substrate with non-infected cells or as useful combinations (syndrome or geographically orientated) for the investigation of serum samples. IgG absorption as preparatory step for the determination of specific antibodies of class IgM: page 60. Cross reactions within the virus family, especially with Flaviviruses, should be taken into consideration since they may cause false-positive results. The infectious agent can be determined by titration of the sample and comparison of titers.
Antibodies against Plasmodium, Rift valley fever virus, Chikungunya virus.
Microplate ELISA: Anti-TBE Virus, Anti-West Nile Virus, Anti-Dengue Virus

Monospecific determination of antibodies against TBE, West Nile and Dengue virus. Serum dilution 1: 100, conjugate class anti-human IgG or IgM, POD-labelled. 1-point calibration, semiquantitative (IgM) or 3-point calibration, quantitative (IgG). Similar incubation conditions and times: All tests can be combined on one and the same microplate. IgM test kit with IgG/RF absorbent in the sample buffer for IgG absorption as preparatory step for the determination of specific IgM antibodies. Available single ELISA: Antigen TBE TBE Vienna WNV Dengue Order No. EI 2661-9601 G or M, avidity, Ab determination in CSF EI 2661-9601-9 G EI 2662-9601 G or M, avidity EI 266b-9601 G or M Order No. (Anti-FSME) EI 2661-9601 G or M Formats page 94 Incubated ELISA Anti-TBE virus, Anti-WNV, AntiDengue virus.
57
EUROIMMUN
BIOCHIP Mosaics for Infectious Serology

(For formats see pages 149-150)
FI 2821-1001-1 G RESPIRATORY TRACT PROFILE 1

IgG V 1. 2. 3. 4. 5. 6. 7. 8. 9. 10. 11. 12. 13. 14. 15. 16. 17. 18. 19. 20. Field A Verification BIOCHIP*** RSV Adenovirus type 3 Influenza virus type A (H1N1) Influenza virus type A (H3N2) Field B Influenza virus type B Parainfluenza virus type 1 Parainfluenza virus type 2 Parainfluenza virus type 3 Field C Parainfluenza virus type 4 Bordetella pertussis** Bordetella parapertussis** Mycoplasma pneumoniae Field D Coxsackie virus type B1 Coxsackie virus type A7 Echo virus type 7 Chlamydia pneumoniae 1 : 10 IgM 1 : 10
FI 2822-1001-1 G EXANTHEMA PROFILE 1

IgG V 1. 2. 3. 4. Field A Verification BIOCHIP*** HHV-6 Rubella virus* Measles virus Mumps virus Field B VZV EBV-CA** EBV-EA Treponema pallidum Field C HSV-1 HSV-2 Coxsackie virus type B1 Coxsackie virus type A9 Field D Echo virus type 7 Borrelia afzelii Borrelia burgdorferi sensu stricto (CH) Borrelia garinii Field E CMV Candida albicans** Candida krusei*,** Candida tropicalis*,** 1 : 10 IgM 1 : 10
1 : 10
1 : 10 5. 6. 7. 8.
1 : 10
1 : 10
1 : 10
1 : 10 9. 10. 11. 12.
1 : 100
1 : 10
1 : 100
1 : 10 13. 14. 15. 16. 1 : 100 17. 18. 19. 20.
1 : 100
1 : 10
Field E 1 : 100 Haemophilus influenzae*,** Klebsiella pneumoniae* Legionella pneumophila serotype 1*,** Legionella pneumophila serotype 12*,**
1 : 100
1 : 100
FI 2823-1001-1 G LYMPHADENITIS PROFILE 1

IgG V 1. 2. 3. 4. 5. 6. 7. 8. 9. 10. 11. 12. 13. 14. 15. 16. 17. 18. 19. 20. Field A Verification BIOCHIP*** HIV-1* HIV-2* HHV-6 Rubella virus* Field B Measles virus Mumps virus Adenovirus type 3 Parainfluenza virus type 1 Field C EBV-CA** EBV-EA Toxoplasma gondii** Treponema pallidum Field D HSV-1 HSV-2 CMV** Coxsackie virus type B5 Field E Coxsackie virus type A9 Bartonella henselae** Chlamydia trachomatis** Chlamydia pneumoniae 1 : 10 IgM 1 : 10
FI 2824-1001-1 G CENTRAL NERVOUS SYSTEM PROFILE 1

IgG V 1. 2. 3. 4. 1 : 10 1 : 10 5. 6. 7. 8. 1 : 10 1 : 10 9. 10. 11. 12. 1 : 100 1 : 10 13. 14. 15. 16. 1 : 100 1 : 10 17. 18. 19. 20. Field A Verification BIOCHIP*** Rubella virus* Measles virus Mumps virus VZV Field B Adenovirus type 3 EBV-CA** Treponema pallidum Toxoplasma gondii** Field C HSV-1 HSV-2 Coxsackie virus type B1 Coxsackie virus type A7 Field D Echo virus type 7 Borrelia afzelii Borrelia burgdorferi sensu stricto (CH) Borrelia garinii Field E CMV Haemophilus influenzae*,** Listeria monocytogenes 1/2a* Listeria monocytogenes 4b* 1 : 10 IgM 1 : 10
1 : 10
1 : 10
1 : 100
1 : 10
1 : 100
1 : 10
1 : 100
1 : 100
* For clinical evaluation the results must be confirmed with a CE approved test. ** For practical reasons the recommended incubation differs from the standard incubation for this substrate. *** Field A contains a verification BIOCHIP. The incubation was performed correctly if the dots show a visible colour reaction. BIOCHIP Mosaics containing fewer substrates can be produced upon request.
58
EUROIMMUN
BIOCHIP Mosaics for Infectious Serology

(For formats see pages 149-150)
FI 2825-1001-1 G MYOCARDITIS PROFILE 1

IgG V 1. 2. 3. 4. 5. 6. 7. 8. 9. 10. 11. 12. 13. 14. 15. 16. Field A Verification BIOCHIP*** Mumps virus Adenovirus type 3 Influenza virus type A (H1N1) Influenza virus type A (H3N2) Field B Influenza virus type B Parainfluenza virus type 1 Parainfluenza virus type 2 Mycoplasma pneumoniae Field C CMV** Coxsackie virus type B1 Coxsackie virus type A16 Echo virus type 7 Field D Borrelia afzelii Borrelia burgdorferi sensu stricto (CH) Borrelia garinii Chlamydia pneumoniae 1 : 10 IgM 1 : 10
FI 2826-1001-1 G INFECTIOUS ARTHRITIS PROFILE 1

IgG V 1. 2. 3. 4. 1 : 10 1 : 10 5. 6. 7. 8. 1 : 100 1 : 10 9. 10. 11. 12. 1 : 100 1 : 10 Field A Verification BIOCHIP*** VZV Influenza virus type A (H1N1) Influenza virus type A (H3N2) Influenza virus type B Field B Yersinia enterocolitica O:3*,** Yersinia enterocolitica O:6*,** Yersinia enterocolitica O:9*,** Toxoplasma gondii** Field C Borrelia afzelii Borrelia burgdorferi sensu stricto (CH) Borrelia garinii Chlamydia trachomatis** 1 : 10 IgM 1 : 10
1 : 10
1 : 10
1 : 100
1 : 10
FI 2827-1001-1 G GASTROINTESTINAL TRACT PROFILE 1

IgG V 1. 2. 3. 4. 5. 6. 7. 8. 9. 10. 11. 12. Field A Verification BIOCHIP*** Adenovirus type 3 Yersinia enterocolitica O:3*,** Yersinia enterocolitica O:6*,** Yersinia enterocolitica O:9*,** Field B Coxsackie virus type B3 Coxsackie virus type A7 Echo virus type 7 Campylobacter jejuni*,** Field C Campylobacter coli*,** Helicobacter pylori** Klebsiella pneumoniae* Candida albicans** 1 : 10 IgM 1 : 10
FI 2828-1001-1 G ACCOMPANYING HEPATITIS PROFILE 1

IgG V 1. 2. 3. 4. Field A Verification BIOCHIP*** Adenovirus type 3 EBV-CA** EBV-EA Toxoplasma gondii** Field B HSV-1 HSV-2 CMV** Coxsackie virus type B5 1 : 10 IgM 1 : 10
1 : 100
1 : 10 5. 6. 7. 8.
1 : 100
1 : 10
1 : 100
1 : 100
Field C 9. Coxsackie virus type A9 10. Echo virus Typ 7 11. Echinococcus granulosus**
1 : 10
1 : 10
FI 2829-1001-1 G OPHTHALMOLOGY PROFILE 1

IgG V 1. 2. 3. 4. 5. 6. 7. 8. Field A Verification BIOCHIP*** Measeles virus VZV Adenovirus type 3 Toxoplasma gondii** Field B HSV-1 HSV-2 Coxsackie virus type A24 Chlamydia trachomatis** 1 : 10 IgM 1 : 10
FI 2831-1001-1 G PREGNANCY PROFILE 1

IgG V 1. 2. 3. 4. 1 : 100 1 : 10 5. 6. 7. 8. Field A Verification BIOCHIP*** Rubella virus* Toxoplasma gondii** HIV-1* HIV-2* Field B Chlamydia trachomatis** CMV** HSV-2 VZV** 1 : 10 IgM 1 : 10
1 : 100
1 : 10
* For clinical evaluation the results must be confirmed with a CE approved test. ** For practical reasons the recommended incubation differs from the standard incubation for this substrate. *** Field A contains a verification BIOCHIP. The incubation was performed correctly if the dots show a visible colour reaction. BIOCHIP Mosaics containing fewer substrates can be produced upon request.
59
EUROIMMUN
Additional Reagents for the Determination of Acute Infections

IgG Absorpion
Before a patients serum is tested for specific antibodies of the IgM class, antibodies of class IgG must be removed by ultracentrifugation, chromatography or immunoabsorption. Specifically bound IgG would displace IgM from the antigen leading to false IgM negative test results. Moreover, the absorption prevents any IgM class rheumatoid factors present from reacting with specifically bound IgG and thus leading to false IgM positive test results. Indication: an IgG absorption of serum samples should always be performed for all IgM antibody determinations in infectious serology before incubating the sera.
rheumatoid factor
IgG
absorbent
EUROSORB IgG/RF Absorbent for Indirect Immunofluorescence

Functional principle: the EUROSORB reagent contains an anti-human IgG antibody preparation from goat. Immunoglobulin G of a serum or plasma sample is bound with high specificity by these antibodies and precipitated. If the sample also contains rheumatoid factors, these will be absorbed by the anti-human IgG/ IgG complex. Incubation time of the sample with the reagent is 15 minutes. A centrifugation step for a subsequent investigation by immunofluorescence is recommended. All IgG subclasses are bound and precipitated by the anti-human IgG antibodies. Human serum IgG in concentrations of up to 15 mg/ml and rheumatoid factors are completely removed by the absorbent (average serum IgG concentration in adults: 12 mg/ml). The recovery rate of the IgM fraction is almost 100%. One unit contains 4.5 ml absorbent, sufficient for the absorption of 100 serum samples.
EUROSORB IgG/RF Absorbent.
Order No. ZF 1270-0145
Formats page 152
Urea Solutions and Avidity Buffers for the Determination of LowAvidity Antibodies in Infectious Serology
To identify low-avidity antibodies in a patients serum, two immunofluorescence tests are performed in parallel: one test is carried out in the conventional way, the other one includes urea treatment between incubations with patients serum and peroxidase-labelled anti-human IgG, resulting in the detachment of lowavidity antibodies from the antigens. Low-avidity antibodies are present if the fluorescence intensity is significantly reduced (two intensity levels ore more) by urea treatment. The following reagents for avidity determination are available: IFT, Ab against Reagents for determination of low-avidity antibodies in infectious serology. Rubella WNV Order No. ZF 1130-0501 ZF 1130-0601 Avidity Solution urea solution, 5 M urea solution, 6 M urea solution, 8 M urea solution, 8 M avidity buffer 1 avidity buffer 2 Incubation Time 10 min 10 min 10 min 30 min 10 min 30 min
Toxoplasma gondii ZF 1130-0801 EBV-EA, EBV-CA ZF 1130-0801 CMV ZF 1131-0101-1 VZV ZF 1131-0101-2
Order No. ZF 1130 und ZF 1131
Formats page 152
60
EUROIMMUN
EUROIMMUN PRODUCTS FOR ALLERGOLOGY
61
EUROIMMUN Products for Allergology
EUROIMMUN
Antibodies of Class IgE against Allergens

EUROLINE: Specific IgE
grasses trees weeds mites animal hair mould spores indicators
Determination of specific IgE in serum. Indication: Clarification of allergic reactions to inhalation allergens, food allergens and cross-reactive allergens (pollen-associated food allergies). Serum dilution: undiluted (or 1 : 10); conjugate class anti-IgE (monoclonal), APlabelled. Calibration: 3 indicator bands on each strip for semiquantitative evaluation. Antibodies against up to 20 allergens per strip can be simultaneously monospecifically detected. EUROLINE profiles with different allergen compositions are available for various test requirements: atopy, inhalation, food and cross reactions. The program EUROLineScan from EUROIMMUN has been developed to enable quantitative evaluation of EUROLINE analyses, facilitate management of data, and provide detailed documentation of results. The incubated EUROLINE test strips are scanned using a flatbed scanner. EUROLineScan recognizes the position of the strips, even if they have been placed inexactly, identifies the bands, and measures their intensity.
Incubated EUROLINE Allergy Profile Inhalation. Order No. (Inhalation) DP 3110-1601 E Formats Page 82
EUROIMMUN Microplate ELISA: Total IgE

Determination of the total IgE concentration in serum. Indications: Differentiation between allergic and intrinsic asthma, between allergic and vasomotor rhinitis, and between atopic and seborrhoic dermatitis. Serum dilution 1 : 10; conjugate class anti-IgE (monoclonal), POD-labelled. One microplate well incubated per patient. 4-point calibration, quantitative. Coating: anti-human IgE, polyclonal. The Total IgE ELISA serves as a screening test for allergy diagnostics and provides an indication for the presence of an allergic reaction.
Incubated ELISA Total IgE.
Order No. EV 3840-9601 E
Formats Page 95
Allercoat 6 Microplate ELISA: Specific IgE

AllercoatTM 6 ELISA. Order No. EP ####-0110 E Formats Page 96-115 Determination of allergen-specific IgE concentrations in serum. Indication: identification of allergic reactions to more than 600 different allergens and allergen mixtures. Serum dilution: undiluted, conjugate class anti-human IgE, AP-labelled. One microplate well per allergen/allergen mixture is incubated for each patient. Calibration: 6-point calibration, quantitative; using reference preparation 2 IRP 75/502 from the WHO. The allergens are coupled to paper rings and can be flexibly configured for each sample according to the analysis. Customized pre-assembled microplates with individual allergy parameters are available on request (Order no.: EP 9901-0101). Orders can be made online with the provided software. Please contact us! A light table and special evaluation software are available to supplement the simple manual Allercoat 6 ELISA procedure. Allercoat 6 ELISA are automatable using the EUROIMMUN Analyzer I and the EUROIMMUN Allercoat software.
62
EUROIMMUN
Antibodies of Class IgE against Allergens

Customized Laboratory Software for Flexible Automation of Allercoat System
Convenient online ordering of individual pre-prepared Allercoat microtiter plates. Integrated light table for manual preparation of ELISA plates and sample distribution. Direct control of photometer, automated evaluation via calibration curve and compilation of results. Fully automated incubation of samples and evaluation of results via connection to the EUROIMMUN Analyzer I. Fully automated administration, documentation and archiving of all data. Simple operation using graphic user commands and clear presentation of the most important information at a glance. Connection to commonly used laboratory systems for convenient transfer of requests and results. Additional security through user administration with individual access rights and confirmation step before results editing. AllercoatTM Software.
Individual equipment with allergen rings
allergen rings
Pipette:
50 l per well
Incubate:
60 min, 37 C
Wash:
4x
;;; ;; ;; ;; ;;; ;; ;; ;; ;; ;; ;; ;; ;; ;; ;; ;; ;; ;; ;; ;;;; ;; ;; ;;

enzyme
undiluted samples
Pipette:
50 l per well
conjugate ;;; ;;; ;;; ;;; ;;; ;;; ;;; ;;;
Incubate:
60 min, 37 C
;;; ;;; ;;; ;;; ;;; ;;; ;;; ;;; ;;; ;;; ;;; ;;; ;;; ;;; ;;; ;;;
Wash:
8x
;;;;; ;; ;; ;; ;; ;; yyy @@@ ;;; yyy @@@ ;;; yy @@ ;; ;; ;; ;; ;;;; @@ ;; yy ;; ;; @@@ ;;; yyy @@@ ;;; yyy @@ ;; ;; ;; ;; ;; ;; @@@ yyy ;;; yyy @@@ yy ;;; @@ ;; yy @@@ ;;; yyy @@@ ;;; yyy @@ ;; yy ;; ;; ;; ;; ;; @@@ ;;; yyy @@@ ;;; yyy @@ ;; @@@ ;;; ;; ;; ;; @@@ ;;; ;; ;;yy @@ ;;
stop solution chromogen-/ substrate sol.
Fitting of allergen rings into microplate wells
Pipette:
100 l per well
Incubate:
30 min, 37 C
Pipette:
100 l per well
Evaluate:
photometric measurement at a wavelength of 405 nm
@@@ ;;; yyy
@@@ yy ;;; yyy @@ ;;

63
EUROIMMUN
General Delivery Conditions

Products from EUROIMMUN AG will be delivered according to the following conditions, as long as no other conditions have been agreed in writing. On issuing an order, the buyer acknowledges the delivery conditions. EUROIMMUN does not accept the sales conditions of the buyer, even when EUROIMMUN has not expressly contradicted them. Contract terms are valid for all future business connection even if they have not been expressly agreed upon again. Orders: Orders to EUROIMMUN can be made in writing (including electronic communication) or verbally. Verbally issued orders first become legally binding for EUROIMMUN with a written confirmation from the orderer or with the delivery of goods. Delivery: EUROIMMUN generally delivers to end users within 7 days of order receipt and to distributors within 14 days, but is not tied to any definite delivery period. If a delivery is not possible through unforseeable circumstances (e.g., factory disruptions, raw material delivery delays, transport difficulties, strikes), the delivery obligation does not apply. EUROIMMUN reserves the right to deliver in part. The delivery obligation of EUROIMMUN AG ceases as long as the customer is in arrears. EUROIMMUN chooses the packing and dispatching method at its own discretion, according to the respective requirements. Product characteristics: The delivered products comply with specifications given in the product catalogue, on the product itself, or in the supplied information sheets. If specifications are inconsistent, the labels on the product itself and the details in the information sheet provided with the product apply. After the given expiration date has passed, the test reagents must not be used. EUROIMMUN products must only be used for the intended purpose. It is the buyers responsibility to check the functional capability immediately on receipt of the product as well as on every day of usage. In particular, the functioning of test reagents can be perturbed through causes outside the direct influence of the maunfacturer, for example, through suboptimal transportation, incorrect storage, or incorrect usage. When test reagents delivered from EUROIMMUN are used, the user must supervise the analysis with appropriate proficiency. Warranty and liability: If nothing else has been agreed upon in writing, all risks pass to the buyer as soon as the goods have been delivered to the carrier or has left EUROIMMUN AG premises for dispatch. With notification of dispatch by EUROIMMUN the risk passes to the buyer if the delivery is postponed or delayed by request of the buyer. On receipt of the goods, the buyer has one working day to check if they are in accordance with the nature and quantity of the agreement and if the goods are free from visible defects. If any complaints arise from this inspection, they should be communicated to EUROIMMUN AG in writing within 2 working days. Hidden defects or functional faults that were not identifiable with the initial inspection and are later discovered should be communicated to EUROIMMUN AG in writing within 14 days of receipt of the goods. If no complaints are received within the stipulated period, it is assumed that the goods are of appropriate quality and quantity for the customer. Complaints do not release the buyer from payment obligation. With prompt and reasonable complaints on the grounds of product defects or the delivery of something other than the ordered goods, EUROIMMUN is obliged to exchange or amend the goods within 14 days or to withdraw or reimbourse the payment, as it chooses. If the defect is not corrected in spite of delivery of a replacement or an amended product, the buyer can demand that the sale is cancelled. If defects are promptly reclaimed, EUROIMMUN has the choice between a further delivery or an appropriate credit note. Further compensatory claims from the buyer are excluded, as long as EUROIMMUN has not violated its contractual or legal obligations through outright negligence or by intention. In such cases EUROIMMUN provides compensation of up to a maximum of 20 times the price of one packet (test kit) of the defective product. EUROIMMUN takes no responsibility for any damage resulting from faulty goods. Prices: Prices from the official EUROIMMUN pricelist in the country of the buyer are applicable. Invoices are provided in the agreed currency. Prices are ex works. Expenses for packing and freight as well as for cooling during transport are added on, including costs for the disposal of packaging material by the customer. Statutory value added tax is not included in the list prices. Payment terms: Payment obligations in countries of export must be settled within 30 days after recieving the goods. No cash discounts will be given unless agreed in writing. Payments by cash transfer or check are valid from the timepoint that the invoice amount is credited to a EUROIMMUN AG bank account. In cases of payment arrears, EUROIMMUN reserves the right to charge compensatory interest of 10% p.a., applicable from the settlement date, without any further notice. In special cases EUROIMMUN can arrange alternative payment periods or demand advance payments. EUROIMMUNs demands resulting from an order cannot be offset by the customer through counter demands. Ownership rights: The delivered goods remain the property of EUROIMMUN AG until full payment. Selling on of EUROIMMUN products is only permitted for companys who are authorized in writing from EUROIMMUN to do so. Software received from EUROIMMUN should not be passed on to third parties without written permission from EUROIMMUN AG. Consultation: Advice from EUROIMMUN AG, although provided to the best knowledge, is nevertheless not binding. No liability claims can ensue from erroneous advice. Applicable law, place of jurisdiction, ineffective regulations: The contract conditions are subject to the laws of the Federal Republic of Germany. The United Nations Conventions on Contracts for the International Sale of Goods does not apply. The place of jurisdiction and fulfilment is Luebeck. If any provision of these general delivery conditions will be held invalid or unenforcable, this general delivery condition will not be rendered invalid as a whole, and the provisions will be changed and interpreted so as best to accomplish the objective of the unenforcable or invalid provision.
153
EUROIMMUN
Index
Autoantibody Diagnostics acetylcholine receptor (ACHR) .............................................................................................. 12, 17, 89 actin ...................................................................................................... 16, 17, 32, 38, 47, 68, 122, 129 adrenal cortex ........................................................................................................... 13, 17, 45, 66, 116 agglutination test ................................................................................................................................ 89 AMA ..................................................... 6, 13, 16-17, 26, 33, 37-40, 45, 47, 68, 79, 116, 122, 126-131 amphiphysin ............................................................................................................ 17, 42, 80, 117-118 ANA ......................................... 14, 16-17, 26, 32-34, 37-38, 42, 46-47, 68, 80, 87, 118-124, 126-131 ANCA ................................................................ 12-14, 16-17, 32, 46-47, 66-67, 86, 119-121, 123-124 aquaporin .................................................................................................................. 12, 17, 42, 66, 118 ASCA ......................................................................................................... 15, 17, 76, 95, 101, 123-124 ASMA ........................................................................ 12, 16-17, 37-38, 68, 117-118, 123-126, 128-131 Autoantibody Profile 2 ...................................................................................................................... 128 basement membrane ...................................................................... 12, 16-17, 46, 67, 80, 86, 120-121 2-glycoprotein ................................................................................................................................... 88 bile ducts ........................................................................................................................................ 17, 32 BP180 ......................................................................................................................... 12, 17, 41, 87, 125 BP230 ......................................................................................................................... 12, 17, 41, 87, 125 BPI ....................................................................................................................................... 17, 46-47, 86 brain ................................................................................................................................ 12, 66, 116-117 C1q ........................................................................................................................................... 16, 87, 88 cANCA ............................................................................... 12, 16-17, 32, 46-47, 66, 86, 119-121, 123 cardiolipin (AMA M1) ....................................................................................................... 16-17, 84, 88 cartilage ........................................................................................................................................ 16, 124 cathepsin G ........................................................................................................................ 17, 46-47, 86 cell nuclei (ANA global test) .... 14, 16-17,26, 32-34, 37-38, 42, 46-47, 68, 80, 87,118-124, 126-131 centromere ................................................................................................ 14, 16, 32, 34, 47, 68, 87-88 cerebellum .................................................................................................... 12-14, 42-44, 83, 116-118 circulating immune complexes (CIC) ................................................................................... 16, 35, 88 cyclic citrullinated peptide (CCP) ........................................................................................ 16, 36, 87 cyclin I (PCNA) ......................................................................................... 12, 16, 26, 32, 47, 68, 80, 87 cyclin II (mitosin) .................................................................................................................... 12, 47, 68 cytoplasm of granulocytes ........................... 12-14, 16-17, 32, 46-47, 66-67, 86, 119-121, 123-124 desmoglein ....................................................................................................... 12, 17, 41, 87, 124, 125 desmosomes ............................................................................................................. 12, 17, 41, 67, 125 dsDNA ...................................... 6, 12, 16-17, 26, 32-35, 38, 42, 47, 68, 79-80, 87, 89, 128, 130, 151 elastase ............................................................................................................................... 17, 46-47, 86 elastin ................................................................................................................................. 12, 16-17, 69 ENA .................................................................................................................. 16, 24, 32, 34, 79-80, 87 endomysium ....................................................................................... 12, 17, 32, 48, 69, 88, 131-133 endothelial cells .................................................................................................. 12, 16-17, 32, 69, 133 epidermis .................................................................................................................. 12-13, 17, 67, 125 EUROArray .......................................................................................................................................... 89 EUROPLUS ............. 7, 12, 32, 37-41, 46-48, 50, 52, 116, 120-123, 126-127, 129, 131-136, 140, 148 eye antigens ....................................................................................................................................... 118 F-actin ...................................................................................................................... 17, 38, 68, 122, 129 GAD ......................................................................................................... 12, 17, 42, 44, 66, 86, 89, 116 gangliosides ................................................................................................................................... 17, 80 gliadin .............................................................................. 7,12, 17, 48, 69, 76, 95, 101, 131-133, 151 glomerular basement membrane (GBM) .......................................... 12, 17, 46, 67, 80, 86, 120-121 glutamate receptor .................................................................................................... 12, 17, 43, 66,118 glutamic acid decarboxylase ............................................................... 12, 17, 42, 44, 66, 86, 89, 116 Golgi apparatus .......................................................................................................... 12, 16, 32, 47, 68 goblet cells ............................................................................................................... 12, 17, 67, 123-124 heart muscle ................................................................................................................ 12, 17, 124, 130 HEp-2-cells ................................................... 6, 12-14, 32-33, 35, 37-38, 44-45, 81, 114-118, 121-126 histones ............................................................................................ 6, 13, 16, 26, 32-34, 47, 79-80, 87 Hu ...................................................................................................... 13, 17, 42-43, 66, 80, 83, 116-118 HUVEC ................................................................................................................................................ 133 hypothalamus antigens ............................................................................................................. 13, 117 IA-2 ........................................................................................................................................... 17, 44, 89 insulin ...................................................................................................................................... 17, 44, 89 intestinal goblet cells .............................................................................................. 12, 17, 67, 123-124 intrinsic factor ............................................................................................ 13, 17, 45, 67, 86, 122-123 Jo-1 .................................................................... 13, 16, 24, 26, 32-34, 47, 68, 79-80, 87-88, 126-127 keratin ............................................................................................................ 12, 13, 16-17, 36, 67, 125 Ku .................................................................................................................... 13, 16, 26, 33, 47, 68, 80 lactoferrin .................................................................................................... 17, 46-47, 67, 86, 121, 124 latex agglutination test ........................................................................................................................ 89 LC-1 ............................................................................................................. 17, 26, 32, 38-39, 79-80, 83 Leydig cells .............................................................................................................. 17, 43, 64, 112-113 liver-kidney microsomes (LKM) ................................................. 13, 17, 37-38, 67, 116, 122, 126-131 liver-specific antigens ........................................................................................ 13, 17, 32, 38, 67, 121 LKM-1 ............................................................................................................... 17, 26, 32, 39, 79-80, 86 lung alveoli .................................................................................................................................. 17, 121 lymphocyte antigens .......................................................................................................... 13, 17, 121 M2 antigen (AMA-M2) .................................................................. 6, 17, 26, 33, 37-39, 68, 79-80, 88 M4 antigen .......................................................................................................... 13, 17, 32, 37, 79, 108 M9 antigen ................................................................................................... 13, 17, 32, 37, 68, 79, 108 Mab .................................................................................................................................. 17, 40, 66, 116 MAG ................................................................................................................................... 13, 17, 42, 66 Mi-2 ........................................................................................................................ 13, 16, 26, 33, 47, 80 mitochondria (AMA) ....................... 6, 13, 16-17, 26, 33, 37-40, 45, 47, 68, 79, 116, 122, 126-131 mitosin (cyclin II) .................................................................................................................... 12, 47, 68 myelin ........................................................................................................................... 12-13, 17, 42, 66 myeloperoxidase (MPO) ........................................................................ 13, 17, 46-47, 79-80, 86, 120 nDNA ....................................... 6, 12, 16-17, 26, 32-35, 38, 42, 47, 68, 79-80, 87, 89, 128, 130, 151 nerves ......................................................................................................... 12-14, 17, 42, 66, 117-118 neuronal autoantibodies ............................................................................................ 17, 42-43, 80, 83 NMDA receptor ......................................................................................................... 12, 17, 43, 66, 118 NMO ....................................................................................................................................... 17, 42, 118 nRNP/Sm .............................................................................. 16, 24, 26, 32-34, 47, 79-80, 87, 126-127 nuclear membrane ........................................................................................................... 17, 32, 47, 68 nucleosomes .............................................................................................. 16, 32-33, 35, 47, 79-80, 87 oesophagus ............................................................................................... 12-13, 41, 48, 125, 131-132 ovarial antigens .............................................................................................................. 45, 86, 89, 116 P/Q calcium channel (VGCC) ....................................................................................................... 17, 89 pANCA ......................................................................... 13, 16-17, 32, 46-47, 67, 86, 119-121, 123-124 pancreas islets ................................................................................................. 114, 44-45, 66, 116, 118 parathyroid gland ................................................................................................................. 13, 17, 116 parietal cells (PCA) ............................................................................. 12, 17, 45, 67, 86, 116, 122-123 parotid gland excretory ducts and acini .......................................................................... 14, 17, 124 PCNA (cyclin I) ......................................................................................... 12, 16, 26, 32, 47, 68, 80, 87 phosphatidylserine ........................................................................................................................ 16, 88 phospholipids .............................................................................................................................. 16, 108 pituitary gland antigens ..................................................................................................... 13, 17, 117 PL-12 ............................................................................................................................ 16, 26, 33, 47, 80 PL-7 .............................................................................................................................. 16, 26, 33, 47, 80 placenta ................................................................................................................................ 14, 17, 117 PM-Scl ..................................................................................................... 6, 16, 26, 32-34, 47, 79-80, 87 proteinase 3 (PR3) ....................................................................... 7, 14, 17, 46-47, 66, 79-80, 86, 120 proteinase 3 (capture test) ........................................................................................................... 46, 86 renal glomeruli (GBM) ......................................................................... 12, 17, 46, 67, 80, 86, 120-121 reticulin ........................................................................................................................... 14, 17, 131-133 retina ................................................................................................................................. 13-14, 17, 118 rheumatoid factors ........................................................................................................................ 36, 60 Ri ............................................................................................................. 17, 42-43, 66, 80, 83, 116-118 RIA ................................................................................................................... 16, 30, 35, 40, 44, 86-90 ribosomal P-proteins ................................................................................................. 14, 16, 32, 68, 88 RNS-Polymerase ............................................................................................................... 14, 16, 32, 47 Ro-52 ......................................................................................... 16-17, 26, 32-34, 39, 47, 79-80, 83, 87 Sa ............................................................................................................................................. 16, 36, 87 Saccharomyces cerevisiae ..................................................................... 15, 17, 76, 95, 101, 123-124 Scl-70 .................................................................. 14, 16, 24, 26, 32-34, 47, 68, 79-80, 87-88, 126-127 skeletal muscle ........................................................................................ 12-14, 17, 38, 122, 124, 130 SLA/LP ........................................................................................................ 17, 26, 32, 38-39, 79-80, 86 Sm ........................................................................... 14, 16, 24, 26, 32-34, 47, 68, 79-80, 87, 126-127 smooth muscles ........................................................................... 12, 16-17, 38, 67, 117-118, 123-126 Sp100 ...................................................................................................................... 17, 26, 38-39, 68, 80 spermatozoa ....................................................................................................... 14, 17, 66, 86, 89, 117 spindle fibers .................................................................................................................... 16, 32, 47, 68 SS-A ......................................................... 6, 14, 16-17, 24, 26, 32-34, 47, 68, 79-80, 83, 87, 126-127 SS-B ........................................................... 6, 14, 16-17, 24, 26, 32-34, 47, 68, 79-80, 87-88, 126-127 ssDNA ..................................................................................................................... 14, 16-17, 34, 47, 87 TAb ................................................................................................................................... 17, 40, 66, 116 testis ............................................................................................................. 14, 17, 40, 43, 66, 116-117 thrombocyte antigens .................................................................................................... 14, 17, 67, 121 thyroglobulin (TG) .................................................................... 14, 17, 40, 45, 66, 79, 86, 89, 90, 116 thyroid gland ......................................................................................... 14, 17, 40, 45, 66, 86, 89, 116 thyroid peroxidase (TPO) .......................................................................................... 17, 40, 79, 86, 89 TRAb .................................................................................................................................. 17, 40, 86, 89 transglutaminase .................................................................................................................... 17, 48, 88 U1-nRNP ............................................................................................................................ 14, 16, 68, 87 vascular endothelium ............................................................................................................ 16, 17, 69 vimentin ............................................................................................................................ 14, 16, 47, 68 Yo ...................................................................................................... 14, 17, 42-43, 66, 80, 83, 116-118 zona pellucida ................................................................................................................... 17, 66, 86, 89 Infectious serology Adenovirus ................................................................................................ 15, 18, 58-59, 74-75, 94,144 Afipia felis ....................................................................................................................................... 15,18 Bartonella ............................................................................................................ 15, 18, 58, 71, 72, 138 Bordetella ..................................................................................................... 15, 18, 58, 70, 81, 91, 134 Borrelia .................................................... 15-18, 21, 28, 50-51, 58-59, 70-71, 77, 81, 84, 91, 135-136 Brucella abortus .................................................................................................................................. 92 Campylobacter .................................................................................................... 15, 18, 59, 70, 91, 134 Candida ....................................................................................................... 15, 18, 58-59, 76, 108, 150 Chikungunya virus ......................................................................................................... 15, 57, 76, 150 Chlamydia .................................................................................. 15-16, 18, 56, 58-59, 71, 92, 137-138 Coxsackie virus ............................................................................................ 15, 18, 58-59, 75, 145-146 Crimean Congo fever virus ................................................................................................. 57, 76, 149 Cytomegalovirus (CMV) ..................................................... 7, 15, 21, 58-60, 73, 81, 85, 93, 142, 151 Dengue virus ............................................................................................. 15, 57, 74, 94, 143-144, 150 EBNA ......................................................................................................... 15, 18, 52-53, 76,81, 95, 148 EBV-CA ........................................................................ 7, 15, 18, 21, 52-53, 58-60, 75-76, 95, 147-148 EBV-EA ................................................................................................ 7, 18, 52, 58-60, 76, 95, 147-148 Echinococcus granulosus ................................................................................ 15, 18, 82, 88, 106, 136 Echo virus ............................................................................................................ 15, 18, 58-59, 75, 146 Epstein-Barr virus capsid ag ......... 7, 15, 18, 21, 52-53, 58-60, 75-76, 78, 81, 85, 95, 147-148, 152 EUROPLUS ............. 7, 12, 32, 37-41, 46-48, 50, 52, 116, 120-123, 126-127, 129, 131-136, 140, 148 EUROSORB IgG/RF absorbens ........................................................................................... 60, 64, 152 Haemophilus influenzae .......................................................................................... 15, 18, 58, 70, 134 Hantavirus ........................................................................................................... 15, 57, 75, 81, 95, 147 Helicobacter pylori .......................................................................... 15, 18, 54, 59, 70, 77, 84, 91, 134 Herpes simplex (HSV) ...................................... 15, 18, 21, 55, 58-59, 73, 78, 81, 85, 92-93, 140-141 HHV-6 ......................................................................................................................... 15, 18, 58, 73, 141 HIV ...................................................................................................... 15, 18, 36, 58-59, 72-73, 140-141 Influenza virus ........................................................................ 15, 58-59, 70, 75, 94-95, 134, 144-145 Japanese encephalitis virus ......................................................................................... 57, 74, 143,150 Klebsiella pneumoniae ....................................................................................... 15, 18, 58-59, 70, 134 Legionella .................................................................................................... 15, 18, 58, 71, 91, 136-137 Leishmania ................................................................................................................ 15, 18, 57, 72, 139 Listeria ....................................................................................................................... 15, 18, 58, 71, 136 Measles virus .................................................................................... 15, 18, 21, 58-59, 64, 73, 93, 142 Mumps virus ....................................................................................... 15, 18, 21, 58-59, 73, 93, 142 Mycoplasma .................................................................................................. 15, 18, 58-59, 72, 92, 139 Parainfluenza virus .............................................................................................. 15, 58-59, 75, 95, 145 Parvovirus ...................................................................................................................................... 36, 93 Plasmodium ........................................................................................................ 7, 15, 18, 57, 140, 150 Reiter spirochetes ............................................................................................................................... 70 Respiratory syncytial virus (RSV) ..................................................................... 15, 18, 58, 74, 94, 144 Rift valley fever virus ........................................................................................................... 57, 74, 149 Rubella virus ..................................................................... 7, 15, 18, 21, 73, 81, 85, 93, 140, 142, 149 Sandfly fever viris ............................................................................................................................... 57 SARS coronavirus ....................................................................................................................... 57, 142 TBE virus ......................................................................................... 15, 18, 21, 48, 55, 72, 90, 138-139 Toxoplasma gondii ..................................................................... 7, 15, 18, 21, 57-60, 72, 81, 92, 140 Treponema pallidum .................................................................................... 15, 18, 58, 70, 77, 84, 91 Ureaplasma urealyticum ............................................................................................... 15, 18, 72, 139 Varicella zoster virus (VZV) ............................................................... 15, 21, 58-60, 73-74, 93-94, 143 VlsE ............................................................................................ 15, 28, 50-51, 70, 81, 84, 91, 135-136 West Nile virus .............................................................................................. 7, 15, 21, 57, 74, 94, 143 Yellow fever virus ........................................................................................................ 57, 74, 143 -144 Yersinia enterocolitica ............................................................... 15-16, 18, 59, 71, 78, 84, 91-92, 137 Allergology animal allergens ...................................................................................................................... 19, 98-99 environmental allergens ...................................................................................................... 19,112-113 food ........................................................................................................................ 19, 62, 82-83, 100-10 further allergens ................................................................................................................................ 110 grasses ..................................................................................................................................... 19, 62,107 herbal and flower pollen ...................................................................................... 19, 62, 102, 114-115 house dust ................................................................................................................................... 19, 108 insects ..................................................................................................... 19, 39, 48, 83, 86, 88, 95, 108 mites ................................................................................................................................ 19, 62, 98, 108 moulds .................................................................................................................................... 19,108-110 parasites ................................................................................................................................... 18-19,110 pharmaceutical drugs ............................................................................................................. 19, 96-97 trees ...................................................................................................................................... 19, 110-111 Automation EUROBlotMaster ...................................................................................................................... 26-27, 29 EUROIMMUN Analyzer ........................................................................................................... 22, 62-63 EUROLineScan ............................................................................................................... 4, 26-29, 51, 62 EUROStar ............................................................................................................................................... 8 instruments ............................................................................................................................... 8, 22, 27
155
EUROIMMUN
IIFT
ELISA
Immunoblots
EUROLabOffice
D-23560 Luebeck (Germany) Seekamp 31 Telephone: +49 451 58550 Fax: +49 451 5855591 euroimmun@euroimmun.de
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TECHNIQUES FOR THE SEROLOGICAL INVESTIGATION OF ANTIBODIES

Techniques for the Serological Investigation of Antibodies

Indirect Immunofluorescence: An Easy and Modern Method

specific human antibody

fine-granular. Pattern homog. Anti- Pattern dsDNA? Anti-Histones? Anti-SS-A? Anti-SS-B?

EUROIMMUNs Innovations for the Standardization and Modernization of Indirect Immunofluorescence

BIOCHIP Technology and Mosaics.

Chemically Activated Cover Glasses for Histochemistry

Fixation of frozen tissue sections to glass surfaces by covalent bonding.

Determination of Low-Avidity Antibodies

low-avide Ab against EBV-CA

high-avide Ab against EBV-CA without urea with urea

EUROPLUS System: Combination of conventional immunofluorescence substrates and monospecific tests

Techniques for the Serological Investigation of Antibodies

Indirect Immunofluorescence: An Easy and Modern Method

Indirect Immunofluorescence: An Easy and Modern Method

Above: conventional slide Below: hydrophobic slide.

Fluorescence Microscope EUROStar II

Techniques for the Serological Investigation of Antibodies

The Indirect Immunofluorescence Test, Performed Using the TITERPLANE Technique

slide BIOCHIPs reagent tray

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1 s flush 5 min cuvette

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1 s flush 5 min cuvette

glycerol/PBS cover glass

Techniques for the Serological Investigation of Antibodies

The Indirect Immunofluorescence Test, Performed Using the TITERPLANE Technique

Recommended Serum Dilutions for Indirect Immunofluorescence Autoimmunity

100 10 + 100 100 10 + 100

10 10 1 10 100 100 100 10

10 10 10 100 100 100

10 100 + 1000 + 10000 10 100 10 100 1 10 10 100 10 10 10 10 10 10

10 10 + 100 10 100 + 1000 + 10000 10 10

10 10 10 10 10 + 100 10 100 + 1000 + 10000 10

100 10 100 10 + 100 10 10 100

10 + 100 100 10 + 100 100

1 100 10 + 100 100 10 10 + 100 10 + 100 10 + 100 100 100 10 1 10 10

Techniques for the Serological Investigation of Antibodies

Recommended Serum Dilutions for Indirect Immunofluorescence Autoimmunity

Recommended Serum Dilutions for Indirect Immunofluorescence Autoimmunity

*) In addition to the preferential analysis or as a plausibility check.

IgA 10 + 100 100

100 100 10 10 10 10 10 + 100

Techniques for the Serological Investigation of Antibodies

Recommended Serum Dilutions for Indirect Immunofluorescence Infectious Serology

Diagnostically Relevant Autoantibodies

CIRC. IMMUNE COMPLEXES C1q ELISA

Grey boxes: standard analysis

*) ANCA diagnostics in acute cases within one hour, at any hour

**) Use special procedure for taking sample(s)

***) Send frozen sample(s)

Diagnostically Relevant Autoantibodies

Organ-/Tissue-Specic Autoimmunity: Autoantibodies against

Antibodies for Infectious Serology

Antibodies for Allergology