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 Title
[Evaluation of the SD Dengue
Duo diagnosis system for
detection of NS1 protein and
IgM and IgG dengue
antibodies].
A custom-designed
recombinant multiepitope
protein as a dengue diagnostic
reagent
A dual marker label free
electrochemical assay for
Flavivirus dengue diagnosis
A rapid point-of-care test for
dengue virus-1 based on a
lateral flow assay with a near-
infrared fluorescent dye.
A systematic review and meta-
analysis of the diagnostic
accuracy of rapid
immunochromatographic
assays for the detection of
dengue virus IgM antibodies
during acute infection
Antibody to the nonstructural
protein NS1 of Japanese
encephalitis virus: Potential
application of mAb-based
indirect ELISA to differentiate
infection from vaccination
Applications of a rapid and
sensitive dengue DUO rapid
immunochromatographic test
kit as a diagnostic strategy
during a dengue type 2
Epidemic in an Urban City
Apropos 'The development of
a novel serotyping-NS1-ELISA
to identify serotypes of
dengue virus'
Abstract Note
INTRODUCTION: SD Dengue Duo (Standard Diagnosis) commercial kit is an immunochromatographic rapid test that detects NS1 protein and IgG/IgM dengue antibodies simultaneously.;
OBJECTIVE: to evaluate the operational and functional characteristics of this system for the detection of virological and serological markers.; METHODS: sera panel was made up by 161
samples; 113 from patients with clinically and serologically confirmed dengue caused by any of the four dengue virus serotypes and 48 negative samples. All these samples were tested by
SD Dengue Duo Kit and by Platelia Dengue NSI Ag; IgM Capture ELISA and ELISA Inhibition Method used as reference assays.; RESULTS: the evaluated kit showed a 57.75% sensitivity for the
detection of NS1 protein; false negatives were detected in samples collected 5 days or more after fever onset in secondary infection cases. IgM detection showed 96.0% sensitivity and
98.4% specificity. Furthermore; high agreement (95.7%) in classifying dengue infection types (primary or secondary infections) was observed. The global study of the 3 markers; the
sensitivity rose to 100%.; CONCLUSIONS: SD Dengue Duo is a simple; easy and rapid assay, it does not require additional equipment; can be used for acute and convalescence serum
samples and offers a good alternative for dengue diagnosis in those laboratories where a complete dengue virus diagnosis is difficult to perform.
Currently; dengue fever is the most important re-emerging mosquito-borne viral disease; with the major proportion of the target population residing in the developing countries of the
world. In endemic areas; potentially fatal secondary dengue infections; characterized by high anti-dengue IgG antibody titers; are most common. Most currently available commercial
dengue diagnostic kits rely on the use of whole virus antigens and are consequently associated with false positives due to serologic cross-reactivity; high cost of antigen production; and
biohazard risk. This has prompted the need to develop an alternate antigen to replace the whole virus antigen in diagnostic tests. We have designed and expressed a novel recombinant
protein antigen by assembling key immunodominant linear IgG-specific dengue virus epitopes; chosen on the basis of pepscan analysis; phage display; and computer predictions. The
recombinant dengue multiepitope protein was expressed to high levels in Escherichia coli; purified in a single step; yielding > 25 mg pure protein per liter culture. We developed an in-house
enzyme-linked immunosorbent assay (ELISA) to detect anti-dengue antibodies in a panel of 20 patient sera using the purified recombinant dengue multiepitope protein as the capture
antigen. The ELISA results were in excellent agreement with those obtained using a commercially available diagnostic test; Dengue Duo rapid strip test from PanBio; Australia. The high
epitope density; careful choice of epitopes; and the use of E. coli system for expression; coupled to simple purification; jointly have the potential to lead to the development of an
inexpensive diagnostic test with a high degree of sensitivity and specificity. (c) 2005 Elsevier Inc. All rights reserved.
Dengue is a RNA viral illness of the genus Flavivirus which can cause; depending on the pervasiveness of the infection; hemorrhagic dengue fever or dengue shock syndrome. Herein we
present an electrochemical label free approach enabling the rapid sensitive quantification of NS1 and IgG (supporting an ability to distinguish primary and secondary infections). Using a
bifunctional SAM containing PEG moieties and a tethered redox thiol; both markers are detectable across clinically relevant levels by label free impedance derived redox capacitance. A
subsequent frequency specific immittance function approach enables assaying (within seconds) with no impairment of analytical quality (linearity; sensitivity and variance). © 2017 Elsevier
B.V.
Dengue fever is caused by the dengue virus (DENV); and DENV1 is the prevalent epidemic serotype in south China. A new lateral flow assay (LFA) based on a near-infrared (NIR) fluorescent
dye was developed to detect anti-DENV1 IgG antibodies. DyLight-800 was used as the marker conjugated to goat anti-human IgG antibodies; and recombinant dengue type 1 envelope
protein was used as the capture protein on the test line. Twenty samples from patients infected with DENV1 and 160 negative controls were analyzed using this new NIR-LFA. The results of
the NIR-LFA were compared with the results of Panbio Dengue IgG ELISA and the Dengue Duo IgM/IgG Cassette. Nineteen confirmed DENV1-positive samples were identified by NIR-LFA;
giving 95% (19/20) sensitivity. No significant differences existed in the results when the 20 primary clinical samples were analyzed using NIR-LFA; Panbio ELISA; and the Dengue Duo
Cassette. However; NIR-LFA had a lower limit of detection than IgG ELISA and Duo IgM/IgG Cassette did when analyzing a 2-fold dilution series of the 19 samples positively identified by NIR-
LFA. When incorporated with an NIR POCT device; the new NIR-LFA was rapid; easy to use; and highly sensitive in detecting DENV1; and has potential for application to clinical
diagnosis.Copyright © 2018 Elsevier B.V. All rights reserved.
A meta-analysis of rapid (≤60 min) dengue diagnostic assays was conducted to determine accuracy and identify causes of between-study heterogeneity. A systematic review identified 302
potentially suitable studies; of which 11 were selected for meta-analysis. All selected studies evaluated the immunochromatographic test (ICT) manufactured by Panbio Pty Ltd. Individual
study results for sensitivity ranged from 0.45 to 1.0; specificity 0.57-1.0; diagnostic odds ratio 4.5-1287; and positive:negative likelihood ratios 2.3-59 and 0.01-0.56; respectively. Results
indicated that the ICT evaluated in the selected studies can both rule in and rule out disease but is more accurate when samples are collected later in the acute phase of infection.
Limitations of this meta-analysis were significant between-study heterogeneity caused by inconsistencies in evaluation methodologies; and the evaluation of only the Panbio ICT. It is
recommended that additional; standardized evaluations are required for other dengue ICTs. © 2005 Royal Society of Tropical Medicine and Hygiene.
An indirect enzyme-linked immunosorbent assay (ELISA) was developed to detect and differentiate the antibody responses to Japanese encephalitis (JE) virus nonstructural protein NS1
between infected and vaccinated individuals. The results showed that all convalescent sera from JE patients contained NS1-specific IgG antibodies; while 65 and 40% of these sera showed
detectable NS1-specific IgM and IgA antibodies; respectively. Specificity analysis showed that NS1-specific IgM and IgA antibodies from JE patients do not cross-react to dengue virus NS1
glycoprotein; while IgG antibodies from 10% of JE patients showed significant cross-reaction to dengue virus NS1 glycoprotein. To differentiate infection from vaccination; the immune sera
from 24 children vaccinated with inactivated JE vaccine were analyzed. The data showed that none of these immune sera had detectable NS1-specific IgG antibodies. The results
demonstrated the potential application of JE NS1-specific indirect ELISA to differentiate infection from vaccination. © 2001 Elsevier Science Ltd.
Dengue infection is a major health problem in tropical and subtropical countries. A prospective observational study in a university-affiliated hospital was conducted between August 2015
and September 2015. Patients who visited the emergency department (ED) with a presentation of any symptoms of dengue were eligible for the dengue non-structural protein 1 (NS1);
IgM/IgG rapid immunochromatographic tests and real-time polymerase chain reaction (RT-PCR) to evaluate the performance of the rapid tests. Considering the RT-PCR as the gold standard
for the dengue diagnosis; the ideal primary results of sensitivity (80-100%); specificity (60-84%); positive predicted value(75%-95%); and negative predicted value (70-100%) suggested that
the NS1-based test with or without a combination of IgM and IgG tests have good diagnostic performances in detecting dengue infections; even in the afebrile or elderly populations. ©
2016 Shih et al. This is an open access article distributed under the terms of the Creative Commons Attribution License; which permits unrestricted use; distribution; and reproduction in
any mediu;M provided the original author and source are credited.

Biological diagnosis of dengue
Clinical and virological factors
influencing the performance of
a ns1 antigen-capture assay
and potential use as a marker
of dengue disease severity
Clinical Evaluation of Dengue
RNA; NS1; and IgM for
Diagnosis of Dengue in
Southern China.
Comparative evaluation of
validity and cost-benefit
analysis of rapid diagnostic
test (RDT) kits in diagnosis of
dengue infection using
composite reference criteria: A
cross-sectional study from
south India
Comparative evaluation of
various commercial assays for
diagnosis of dengue fever.
Comparison of a rapid
immuno-chromatography
assay with a standard ELISA
for the detection of IgM and
IgG antibodies against dengue
viruses
Comparison of five serological
diagnostic assays for the
detection of IgM and IgG
antibodies to dengue virus
Comparison of PanBio dengue
duo enzyme-linked
immunosorbent assay (ELISA)
Dengue is the most important disease caused by an arbovirus worldwide. Its clinical manifestations are very large from asymptomatic infections to severe diseases with fatal outcome. No
effective antiviral treatment or vaccine is available. Thus; a rapid and accurate diagnosis is of paramount importance both for better clinical case management and surveillance. Diagnosis
methods depend on the time clinical signs appeared.Within the 7 first days of fever; direct tests are preferred. RT-PCR methods are sensitive; specific; and can identify viral serotypes.
Conventional RT-PCR will probably be replaced by real time PCR as soon as standardised and accurate assays for the four serotypes will be available. Serology (EIA) is used only after 7 days
of disease; i.e. late in the course of dengue, it is accurate; specific but not discriminatory for serotypes and high cross-reactive. NS1 antigen detection still lack of clinical sensitivity and viral
isolation is too fastidious. Even though ameliorations are necessary; viral detection by RT-PCR remains the best tool in clinical settings for a rapid diagnosis of severe dengue infections.
Background: Detection of dengue NS1 antigen in acute infection has been proposed for early diagnosis of dengue disease. The aim of this study was to evaluate the clinical and virological
factors influencing the performance of the Platelia NS1 Ag kit (BioRad) and to assess the potential use of NS1 antigen and dengue viral loads as markers of dengue disease severity.
Methodology/Principal Findings: Blood specimens were collected from patients hospitalized at the Kampong Cham hospital during the 2006 and 2007 dengue epidemics in Cambodia.
Dengue infection was confirmed in 243/339 symptomatic patients and in 17 asymptomatic individuals out of 214 household members tested. Overall sensitivity and specificity of Platelia
NS1 Ag kit were 57.5% and 100% respectively. NS1 Ag assay combined with IgM antibody capture ELISA significantly increased the sensitivity for dengue diagnosis. NS1 Ag positivity rate
was found significantly higher in DF than in DHF/DSS; in primary than in secondary infections; in patients with a high viremia (>5 log/mL) and in patients infected with DENV-1. In
asymptomatic individuals; the NS1 Ag capture sensitivity tends to be lower than that in symptomatic patients. Milder disease severity was observed independently in patients with RNA
copy number >5 log10 cDNA equivalents/mL or in high level of NS1 antigen ratio or in DENV-1 infection. Conclusions: Overall sensitivity of NS1 Ag detection kit varied widely across the
various forms of dengue infection or disease. Sensitivity was highest in patients sampled during the first 3 days after onset of fever; in patients with primary infection; DENV-1 infection;
with high level of viremia and in DF rather than DHF/DSS. In asymptomatic patients; RT-PCR assay has proved to be more sensitive than NS1 antigen detection. The NS1 antigen level
correlated significantly with viremia and a low NS1 antigen ratio was associated with more severe disease. © 2011 Duong et al.
: In 2014; a large outbreak of dengue occurred in Guangzhou; China. This outbreak prompted us to evaluate NS1 and RNA for the early diagnosis of acute dengue infection; in addition to
the combination with IgM antibody. We aimed to find the differences of three assays about dengue diagnosis. This study was an evaluation of diagnosis test. Based on WHO criteria 2009;
dengue RNA; NS1; and IgM/IgG were detected from 294 patients (180 dengue patients; 114 non-dengue patients) by three diagnostic kits made in China. The [chi]2 test; sensitivity; and
specificity were used in statistical analysis. The ratios of dengue patients with low platelet counts (<100 x 109/L 32.2%) or white blood cell counts (<4.0 x 109/L 58.9%) were significantly
higher compared to non-dengue patients (P < 0.05). Dengue NS1 was shown sensitive (93.9%) for diagnostic use. RNA had a better performance with 98.1% of sensitivity from day 1 to day
4 after illness onset. IgM performed better at day 5 or more with 74.0% of sensitivity. The diagnostic rate using a combination of RNA and IgM was 97.8% and 96.7% using NS1 and IgM. A
patient with low platelet and white blood cell counts needs additional tests for dengue during an epidemic. RNA and NS1 were most valuable for early diagnosis of dengue; whereas IgM
was best suited as a supplementary method for patients at day 5 or more after illness onset.; (C) 2016 John Wiley & Sons; Inc
Background & objectives: Rapid diagnostic test (RDT) kits are widely used in India for the diagnosis of dengue infection. It is important to evaluate the validity and reliability of these RDTs.
The study was aimed to determine the sensitivity; specificity and predictive value of four commercially available RDTs [Panbio Dengue Duo cassette; Standard Diagnostics (SD) Bioline
Dengue Duo; J. Mitra Dengue Day-1 test and Reckon Dengue IgG/IgM] against composite reference criteria (CRC); and compare the cost of the tests. Methods: In this prospective
observational study for diagnostic accuracy; we tested stored blood samples from 132 cases of dengue and 149 controls of other infections as classified based on CRC; with all the four
RDTs. The CRC was based on the epidemiological considerations; common clinical features and laboratory abnormalities. The non-dengue controls were the cases of proven alternative
diagnosis. The diagnostic performances of the tests were compared in terms of sensitivity; specificity and predictive value along with the cost involved per test. Results: The sensitivity of
the Panbio and SD RDT kits was found to be 97.7 and 64.3% respectively; and the specificities were 87.8 and 96.6% respectively. The sensitivity of the NS1 antigen capture by SD Duo;
Reckon; J. Mitra RDTs was 20.9; 18.6 and 27.1% respectively. The prevalence of dengue specific IgG antibody with Panbio RDT kits was 49.3%. The cost per test for Panbio; SD; Reckon and J.
Mitra is US$ 6.90; 4.27; 3.29 and 3.61 respectively. Conclusion: It was concluded that in dengue outbreak; Panbio IgM capture RDT alone is reliable and easily available test which can be
used in acute phase of dengue infection in any resource limited set up. NS1 capture rates by any of the other three RDTs might not be reliable for the diagnosis of acute dengue infection. ©
2016; Malaria Research Center. All rights reserved.
Dengue fever (DF) is endemic in India and dengue hemorrhagic fever (DHF) has been reported with increasing frequency in the last decade. We evaluated three commercial assays for
detection of antibodies to dengue virus; to assess their performance in a diagnostic laboratory. Sera from 58 patients collected during a febrile outbreak in New Delhi in 1997 were studied.
The methods evaluated were MRL Diagnostic Dengue Fever Virus IgM Capture ELISA; Pan Bio Dengue Duo IgM and IgG Capture ELISA and Pan Bio Rapid Immunochromatographic test. The
MRL ELISA correctly identified 97.8% (43 of 44) of samples as dengue positive while the Pan Bio Duo ELISA and Pan Bio RIT identified 95.45% (42 of 44). The sensitivities of both Pan Bio Duo
ELISA and Pan Bio RIT for primary dengue and secondary dengue were 100% and 93.54% respectively. The specificity of three assays were MRL IgM ELISA 100%; Pan Bio Duo ELISA 92.8%
and Pan Bio RIT 85.7%.
A total of 765 blood samples collected from dengue suspected patients admitted to a Teaching Hospital in Sri Lanka were used to compare a rapid ICT assay with a standard ELISA for the
detection of anti-dengue virus (DENV) IgM and IgG. Detection accuracy indices including sensitivity; specificity; positive predictive value (PPV); negative predictive value (NPV); Chi square
and Cohen’s kappa values were determined for the ICT assay using the ELISA as a comparator for the detection of anti-DENV IgM and IgG. The rapid ICT assay showed a sensitivity of 64%;
specificity of 75%; NPV of 68% and PPV of 72% for anti-DENV IgM detection. However; all the accuracy indices were relatively higher for the anti-DENV IgG detection by the ICT assay than
those for anti-DENV IgM detection. Despite the low sensitivity for anti-DENV IgM detection by the ICT assay; considering the limitations in using ELISAs in resource limited regions; rapid ICT
assays would be useful for the detection of more recent DENV infections. As many patients present after fever days 5 in the study area; anti-DENV IgM/IgG would be the suitable marker to
be detected by rapid ICT assays in such areas. © 2018; Indian Virological Society.
Difficulties in the management of dengue infections include the lack of rapid diagnostic methods and symptoms of dengue are often confused with those of other diseases. Five commercial
kits were evaluated for the detection of dengue-specific IgM and IgG antibodies using sera obtained from patients with primary and secondary dengue infections; as well as other febrile
illnesses. All kits were compared to the in-house IgM ELISA and HI assays. The rapid test kits took either 30 to 45 min (PanBio Dengue Duo Cassette; Accusens Dengue Virus Rapid Strip Test;
and Unitest Dengue IgM and IgG Combo Rapid Test) or between 2 to 4 h (PanBio Dengue Duo Capture ELISA; and Antivirus IgM Detecion Kit 96 [Pentax Corporation]). Most kits were able to
detect IgM in more than 90% of the secondary convalescent sera; while IgG detection was generally high (80 to 100%). All five kits showed high specificity when tested against sera from
other febrile patients; and have been shown to be extremely useful in the diagnosis of dengue infections.
The performances of the MRL dengue fever virus immunoglobulin M (IgM) capture enzyme-linked immunosorbent assay (ELISA) and the PanBio Dengue Duo IgM capture and IgG capture
ELISA were compared. Eighty sera from patients with dengue virus infections; 24 sera from patients with Japanese encephalitis (JE); and 78 sera from patients with nonflavivirus infections;
such as malaria; typhoid; leptospirosis; and scrub typhus; were used. The MRL test showed superior sensitivity for dengue virus infections (94 versus 89%); while the PanBio test showed
 and MRL dengue fever virus
immunoglobulin M capture
ELISA for diagnosis of dengue
virus infections in Southeast
Asia.
Comparison of PanBio Dengue
Duo Igm and IgG Capture ELISA
and Venture Technologies
Dengue IgM and IgG Dot Blot
Comparison of real-time SYBR
green dengue assay with real-
time taqman RT-PCR dengue
assay and the conventional
nested PCR for diagnosis of
primary and secondary dengue
infection
Comparison of seven
commercial antigen and
antibody enzyme-linked
immunosorbent assays for
detection of acute dengue
infection
Comparison of the diagnostic
accuracy of commercial NS1-
based diagnostic tests for early
dengue infection
Comparison of two dengue
NS1 rapid tests for sensitivity;
specificity and relationship to
viraemia and antibody
responses
Comparison of two rapid
diagnostic tests during a large
dengue virus serotype 3
outbreak in the Solomon
Islands in 2013
superior specificity for JE (79 versus 25%) and other infections (100 versus 91%). The PanBio ELISA showed better overall performance; as assessed by the sum of sensitivity and specificity
(F value). When dengue virus and nonflavivirus infections were compared; F values of 189 and 185 were obtained for the PanBio and MRL tests; respectively; while when dengue virus
infections and JE were compared; F values of 168 and 119 were obtained. The results obtained with individual sera in the PanBio and MRL IgM ELISAs showed good correlation; but this
analysis revealed that the cutoff value of the MRL test was set well below that of the PanBio test. Comparing the sensitivity and specificity of the tests at different cutoff values (receiver-
operator analysis) revealed that the MRL and PanBio IgM ELISAs performed similarly in distinguishing dengue virus from nonflavivirus infections; although the PanBio IgM ELISA showed
significantly better distinction between dengue virus infections and JE. The implications of these findings for the laboratory diagnosis of dengue are discussed.
Background: A number of commercial ELISA for dengue diagnosis have recently become available; though direct comparison between these assays have not been published. Objectives: The
Venture Technologies Dengue IgM and IgG Dot Blot assays and the PanBio Dengue Duo IgM and IgG Capture ELISA were compared. Study design: Paired sera from patients with dengue
(n=20) and Japanese encephalitis (JE; n=10); and single sera from patients with typhoid (n=10); leptospirosis (n=10) and scrub typhus (n=10) were assayed according to the manufacturer's
instructions. Results: The Dot Blot IgM ELISA showed higher sensitivity than the PanBio IgM ELISA (100 vs. 95%); while the PanBio IgM ELISA showed higher specificity in JE (100 vs. 20%) and
non-flavivirus infections (100 vs. 97%). Defining elevation of either IgM or IgG as a positive result; the Dot Blot and ELISA tests both showed 100% sensitivity in dengue infection; while the
PanBio test showed superior specificity in JE (70 vs. 0%) and non-flavivirus infections (100 vs. 67%). Conclusions: Both assays are useful aids to the serological diagnosis of dengue infection.
The clinical setting; user preference and local conditions will be important in determining which test is more appropriate. Copyright (C) 2000 Elsevier Science B.V.
Background: Dengue fever and dengue hemorrhagic fever are caused by dengue virus. Dengue infection remains a burning problem of many countries. To diagnose acute dengue in the
early phase we improve the low cost; rapid SYBR green real time assay and compared the sensitivity and specificity with real time Taqman® assay and conventional nested PCR assay. Aims:
To develop low cost; rapid and reliable real time SYBR green diagnostic dengue assay and compare with Taqman real-time assay and conventional nested PCR (modified Lanciotti). Materials
and Methods: Eight cultured virus strains were diluted in tenth dilution down to undetectable level by the PCR to optimize the primer; temperature (annealing; and extension and to detect
the limit of detection of the assay. Hundred and ninety three ELISA and PCR proved dengue clinical samples were tested with real time SYBR® Green assay; real time Taqman® assay to
compare the sensitivity and specificity. Results: Sensitivity and specificity of real time SYBR® green dengue assay (84% and 66%; respectively) was almost comparable to those (81% and
74%) of Taqman real time PCR dengue assay. Real time SYBR® green RT-PCR was equally sensitive in primary and secondary infection while real time Taqman was less sensitive in the
secondary infection. Sensitivity of real time Taqman on DENV3 (87%) was equal to SYBR green real time PCR dengue assay. Conclusion: We developed low cost rapid diagnostic SYBR green
dengue assay. Further study is needed to make duplex primer assay for the serotyping of dengue virus.
Seven commercial assays were evaluated to determine their suitability for the diagnosis of acute dengue infection: (i) the Panbio dengue virus Pan-E NS1 early enzyme-linked
immunosorbent assay (ELISA); second generation (Alere; Australia), (ii) the Panbio dengue virus IgM capture ELISA (Alere; Australia), (iii) the Panbio dengue virus IgG capture ELISA (Alere;
Australia), (iv) the Standard Diagnostics dengue virus NS1 antigen ELISA (Standard Diagnostics; South Korea), (v) the Standard Diagnostics dengue virus IgM ELISA (Standard Diagnostics;
South Korea), (vi) the Standard Diagnostics dengue virus IgG ELISA (Standard Diagnostics; South Korea), and (vii) the Platelia NS1 antigen ELISA (Bio-Rad; France). Samples from 239 Thai
patients confirmed to be dengue virus positive and 98 Sri Lankan patients negative for dengue virus infection were tested. The sensitivities and specificities of the NS1 antigen ELISAs ranged
from 45 to 57% and 93 to 100% and those of the IgM antibody ELISAs ranged from 85 to 89% and 88 to 100%; respectively. Combining the NS1 antigen and IgM antibody results from the
Standard Diagnostics ELISAs gave the best compromise between sensitivity and specificity (87 and 96%; respectively); as well as providing the best sensitivity for patients presenting at
different times after fever onset. The Panbio IgG capture ELISA correctly classified 67% of secondary dengue infection cases. This study provides strong evidence of the value of combining
dengue virus antigen- and antibody-based test results in the ELISA format for the diagnosis of acute dengue infection. Copyright © 2012; American Society for Microbiology.
Background: We compared the diagnostic accuracy and reproducibility of commercially available NS1-based dengue tests and explored factors influencing their sensitivities. Methods:
Paired analysis of 310 samples previously characterized as positive (n = 218) and negative (n = 92) for viral isolation and/or RT-PCR and/or IgM seroconversion. Masked samples were tested
by two observers with Platelia (TM) Dengue NS1 Ag; second generation Pan-E (TM) Dengue Early ELISA; SD Dengue NS1 Ag ELISA; Dengue NS1 Ag STRIP (TM); and SD BIOLINE (TM) Dengue
Duo (NS1/IgM/IgG). Results: SD BIOLINE (TM) NS1/IgM/IgG had the highest sensitivity (80.7% 95% CI 75-85.7) with likelihood ratios of 7.4 (95% CI 4.1-13.8) and 0.21 (95% CI 0.16-0.28). The
ELISA-format tests showed comparable sensitivities, all below 75%. STRIP (TM) and SD NS1 had even lower sensitivities (<65%). The sensitivities significantly decreased in samples taken
after 3 days of fever onset; in secondary infections; viral serotypes 2 and 4; and severe dengue. Adding IgM or IgG to SD NS1 increased its sensitivity in all these situations. Conclusions: The
simultaneous detection of NS1/IgM/IgG would be potentially useful for dengue diagnosis in both endemic and non endemic areas. A negative result does not rule out dengue. Further
studies are required to assess the performance and impact of early laboratory diagnosis of dengue in the routine clinical setting.
Background: Dengue is a major public health problem in tropical and subtropical countries. Rapid and easy diagnosis of dengue can assist patient triage and care-management. The
detection of DENV NS1 on rapid lateral flow tests offers a fast route to a presumptive dengue diagnosis but careful evaluations are urgently needed as more and more people use them.
Methods: The sensitivity and specificity of the Bio-Rad NS1 Ag Strip and SD Dengue Duo (NS1/IgM/IgG) lateral flow rapid tests were evaluated in a panel of plasma samples from 245
Vietnamese patients with RT-PCR confirmed dengue and 47 with other febrile illnesses. Results: The NS1 rapid tests had similar diagnostic sensitivities (respectively 61.6% and 62.4%) in
confirmed dengue cases but were 100% specific. When IgM/IgG results from the SD Dengue Duo were included in the test interpretation; the sensitivity improved significantly from 62.4%
with NS1 alone to 75.5% when NS1 and/or IgM was positive and 83.7% when NS1 and/or IgM and/or IgG was positive. Both NS1 assays were significantly more sensitive for primary than
secondary dengue. NS1 positivity was associated with the underlying viraemia as NS1-positive samples had a significantly higher viraemia than NS1-negative samples. Conclusions: These
data suggest that the NS1 test component of these assays are highly specific and have similar levels of sensitivity. The IgM parameter in the SD Duo test improved overall test sensitivity
without compromising specificity. The SD Dengue Duo lateral flow rapid test deserves further prospective evaluation in dengue endemic settings.
Dengue virus (DENV) infection causes various clinical presentations; including asymptomatic infection; dengue with or without warning signs and severe dengue. An early and accurate
diagnosis of DENV infection during the first few days of illness supports clinical management and significantly reduces dengue-associated mortality and morbidity. However; it is very
difficult to confirm DENV infection in endemic regions without qualified dengue diagnostic laboratories. In this study; we evaluated the performance of two commercially available rapid
diagnostic tests (RDTs) using serum samples collected in the Solomon Islands during the 2013 DENV-3 outbreak. The sensitivity and specificity of the tests were calculated by comparing the
results of DENV nonstructural protein 1 (NS1); IgM and IgG RDTs with those obtained by qRT-PCR. We also compared the results of the DENV IgM/IgG RDT with those obtained using an
IgM/IgG capture enzyme-linked immune-sorbent assay (ELISA). The sensitivities of the SD and CTK NS1 RDTs were similar (90.9% and 92.6%); and the specificity of the SD NS1 RDT was
significantly higher than that of the CTK NS1 RDT (100% versus 78.8%). The inclusion of IgM and IgG in the RDT did not significantly increase the sensitivity for DENV diagnosis. Compared
with the SD IgM RDT; IgM capture ELISA had the same specificity but higher sensitivity. Userfriendly RDTs remain the first choice and the most convenient tool in dengue endemic regions;
where laboratory facilities and the corresponding infrastructure are lacking. Our study provided important and practical information for comparing the performance and validity of the
different RDTs for rapid dengue detection. © 2018 Liu et al. This is an open access article distributed under the terms of the Creative Commons Attribution License; which permits

Dengue diagnosis; advances
and challenges
Dengue Duo Rapid Test: A
Valuable Screening Test for
Dengue Outbreak
Dengue fever: Its laboratory
diagnosis; with special
emphasis on IgM detection
Dengue in travellers:
applicability of the 1975-1997
and the 2009 WHO
classification system of dengue
fever.
Dengue viremia in blood
donors from Honduras; Brazil;
and Australia.
Dengue virus 4 (DENV-4) re-
emerges after 30 years in
Brazil: Cocirculation of DENV-
2; DENV-3; and DENV-4 in
Bahia
Detection of specific
antibodies in saliva during
dengue infection.
Diagnosing Dengue at the
Point-of-Care: Utility of a
Rapid Combined Diagnostic Kit
in Singapore
unrestricted use; distribution; and reproduction in any mediu;M provided the original author and source are credited.
Dengue diagnosis was one of the topics discussed at the symposium 'The Global Threat of Dengue - Desperately Seeking Solutions' organized during the 10th International Congress of
Infectious Diseases held in Singapore in 2002. In this paper; a review is presented focusing on the main advances; problems and challenges of dengue diagnosis. IgM capture ELISA; virus
isolation in mosquito cell lines and live mosquitoes; dengue specific monoclonal antibodies and PCR have all represented major advances in dengue diagnosis. However; an appropriate
rapid; early and accessible diagnostic method useful both for epidemiological surveillance and clinical diagnosis is still needed. Also; tools that suggest a prognosis allowing for better
management are also needed. Finally; laboratory infrastructure; technical expertise and research capacity must be improved in endemic countries in order to positively influence dengue
surveillance; clinical case management and the development of new approaches to dengue control. © 2003 International Society for Infectious Diseases. Published by Elsevier Ltd. All rights
reserved.
Dengue is a major health problem in many parts of the tropical world presenting as both epidemic & endemic forms. It is a mosquito borne illness caused by one of the serotypes of dengue
viruses. Mortality rate of dengue in untreated patients is 10%-12%. The present study aims to determine whether the rapid test SD dengue duo (IgG; Ig;M and NS-1 Ag detection) can be
used as screening test in dengue outbreak when compared to ELISA. 226 serum samples were tested in patients clinically suspected Dengue. All the 226 samples were subjected to IgG; IgM
Microlisa test. The same were put on rapid SD bioline Dengue duo rapid test and was compared with ELISA. 150 samples were tested positive with ELISA (either positive for IgG; IgM). When
compared with ELISA; Rapid test showed sensitivity of 80.6% specificity and positive predictive value of 100% & zero false positive rates. Efficiency of the test was 87.16% SD Dengue duo
rapid test should be a valuable screening test for dengue fever outbreak which can be interpreted easily. Results were comparable to ELISA. It provides additional diagnostic investigation of
primary and secondary dengue that compliments NS-1 antigen detection.
Rapid diagnosis of dengue is crucial for proper patient care. As IgM antibody appears early during the disease course; its detection is a valuable tool for rapid diagnosis. We evaluated and
compared two commercial tests; the PanBio Rapid Immunochromatographic Card Test (Brisbane; Australia) and the PanBio Microwell IgM ELISA with an IgM Capture ELISA (National
Institute of Virology; Pune; India). A total of 154 samples from individuals with febrile illness having dengue fever (DF)-like symptoms were tested. The NIV IgM Capture ELISA (MAC-ELISA)
showed a high positivity rate (38.9%) as compared to the PanBio Rapid (22.7%) and the PanBio IgM ELISA (20.7%). The NIV MAC-ELISA showed a high sensitivity (96%) as compared to
PanBio Rapid (73%) and PanBio IgM ELISA (72%). But the specificity was low (81%) when compared to PanBio Rapid (95%) and PanBio IgM ELISA (97%) using the latent class analysis model.
The MAC-ELISA; though a three-day procedure; should be a valuable screening test because of its high sensitivity rates. But rapidity gets compromised; as batch testing is required along
with technical difficulty in performance. The "Rapid" test is an easier option in small peripheral laboratories in India because of its obvious advantages.
OBJECTIVES: The aim of this study was to assess the applicability and benefits of the new WHO dengue fever guidelines in clinical practice; for returning travellers.; METHODS: We
compared differences in specificity and sensitivity between the old and the new guidelines for diagnosing dengue and assessed the usefulness in predicting the clinical course of the
disease. Also; we investigated whether hypertension; diabetes or allergies; ethnicity or high age influenced the course of disease.; RESULTS: In our setting; the old classification; compared
with the new; had a marginally higher sensitivity for diagnosing dengue. The new classification had a slightly higher specificity and was less rigid. Patients with dengue who had warning
signs as postulated in the new classification were admitted more often than those who had no warning signs (RR; 8.09 [1.80-35.48]). We did not find ethnicity; age; hypertension; diabetes
mellitus or allergies to be predictive of the clinical course.; CONCLUSIONS: In our cohort of returned travellers; the new classification system did not differ in sensitivity and specificity from
the old system to a clinically relevant degree. The guidelines did not improve identification of severe disease.; Copyright (C) 2012 Blackwell Publishing Ltd.
BACKGROUND: Dengue fever and hemorrhagic disease are caused by four dengue virus (DENV) serotypes (DENV-1 to -4); mosquito-borne flaviviruses with increasing incidence; and
expanding global distributions. Documented transfusion transmission of West Nile virus raised concern regarding transfusion-transmitted DENV.; METHODS: A DENV RNA assay was
developed based on transcription-mediated amplification (TMA) blood screening assays routinely used by blood centers worldwide. Sensitivity was established by endpoint dilution
analyses of DENV-1 RNA transcript and pedigreed tissue culture standards for all four DENV-serotypes. Frozen plasma samples were tested from 2994 donations from Honduras (September
2004-January 2005); 4858 donations from Brazil (February-April 2003); and 5879 donations from Australia (March-September 2003). Type-specific polymerase chain reaction (PCR) assays
were used to quantify and genotype TMA repeat-reactive samples, viral cultures; type-specific antibody; and antigen assays were also performed.; RESULTS: The TMA assay detected 14.9
copies per mL DENV-1 transcript (95% detection limit); with comparable sensitivity for all four serotypes. Honduran donors yielded 9 TMA repeat-reactive samples (0.30%), 8 were
confirmed by PCR; with 3 DENV serotypes detected and viral loads from fewer than 3 x 104 to 4.2 x 104 copies per mL, and 4 samples yielded infectious virus. Three (0.06%) Brazilian
samples tested repeat-reactive, 2 (0.04%) were PCR-positive (serotypes DENV-1 and -3, 12 and 294 copies/mL). No Australian donor samples tested repeat-reactive.; CONCLUSION: Dengue
viremia rates among asymptomatic blood donors ranged from 0.30 percent in Honduras to 0.04 percent in Brazil. Future studies are needed to establish rates of transfusion transmission by
viremic donations and clinical consequences in recipients.; Copyright (C) 2008 Blackwell Publishing Ltd.
Dengue fever (DF) is a mosquito-borne viral disease of great concern in tropical and subtropical regions of the world. One important cause of the increase in DF is rapid development and
urbanization has led to proliferation of the Aedes aegypti mosquito; the vector responsible for transmission of the illness. Surveillance of dengue virus (DENV) infection in Brazil shows the
predominance of DENV-1; DENV-2; and DENV-3 until 2010. This study reports the reappearance of DENV-4 in Brazil for the first time in 30 years. Serum samples were collected from
individuals (n = 214) exhibiting fever and muscular pain in Bahia; Brazil; during 2011–2012. These samples were subjected to reverse transcription-polymerase chain reaction (RT-
PCR)/nested PCR; which revealed that 82% of samples were positive for DENV-4, most were older age groups and exhibited a serological pattern consistent with a primary infection. The
cocirculation of multiple DENV serotypes within the same city places the population at risk for a fatal form of the disease. Therefore; with the increasing incidence of severe DF cases; early
diagnosis will be a priority for public health efforts in Brazil. © 2015; National Institute of Health. All rights reserved.
Saliva was collected prospectively from patients presenting with suspected dengue infection 4 to 8 days after the onset of symptoms and assayed by a commercial dengue immunoglobulin
M (IgM) and IgG capture enzyme-linked immunosorbent assay (ELISA) (PanBio Dengue Duo ELISA). Laboratory diagnosis was based on virus isolation and on hemagglutination inhibition
(HAI) assay and an in-house IgM and IgG capture ELISA. With a positive result defined as either salivary IgM or IgG levels above the cutoff value; an overall sensitivity of 92% was obtained
for both primary- and secondary-dengue patients (22 of 24); while no patients with non-flavivirus infections (n = 11) and no healthy laboratory donors (n = 17) showed elevation of salivary
antidengue antibody (100% specificity). Salivary IgG levels correlated well with serum HAI titer (r = 0.78); and salivary IgG levels could be used to distinguish between primary- and
secondary-dengue virus infections.
WHO recommendations for dengue diagnosis require laboratory facilities. Antibody-based rapid diagnostic tests (RDTs) have performed poorly; and clinical diagnosis remains the mainstay
in dengue-endemic countries. We evaluated a combination antigen-antibody RDT for point-of-care testing in a high-prevalence setting. In this prospective cohort study; adults were
enrolled from a tertiary infectious disease centre for evaluation of undifferentiated febrile illness from October 2011 to May 2012. SD Bioline Dengue Duo was evaluated at point-of-care
against a WHO-based reference standard of viral isolation; RT-PCR; NS1-; IgM-; and IgG-ELISA. 246 adults were enrolled (median age 34 years; range 18-69); of which 197 could be

Diagnosis of Dengue Virus
Infection by the Visual and
Simple AuBioDOT
Immunoglobulin M Capture
System
Differentiating secondary from
primary dengue using IgG to
IgM ratio in early dengue: An
observational hospital based
clinico-serological study from
North India
Distinguishing dengue fever
from other infections on the
basis of simple clinical and
laboratory features:
Application of logistic
regression analysis
Early dengue diagnosis by
nonstructural protein 1
antigen detection: Rapid
immunochromotography
versus two the enzyme-linked
immunosorbent assay kits
Early diagnosis of Dengue
infection using a commercial
Dengue Duo rapid test kit for
the detection of NS1; IG;M and
IGG.
Enhanced performance of an
innovative dengue IgG/IgM
rapid diagnostic test using an
anti-dengue EDI monoclonal
antibody and dengue virus
antigen
Evaluación del sistema
diagnóstico SD Dengue Duo
para la detección de la
proteína NS1 y los anticuerpos
IgM e IgG anti-dengue
confirmed definitively as either dengue or non-dengue. DENV-2 was the predominant serotype (79.5%) and the ratio of primary to secondary cases was 1:1.1. There were no test failures
and minimal interobserver variation with a Fleiss' kappa of 0.983 (95% CI 0.827-1.00). Overall sensitivity and specificity were 93.9% (95% CI 88.8-96.8%) and 92.0% (95% CI 81.2-96.9%)
respectively. Using WHO clinical criteria alone for diagnosis had similar sensitivities (95.9%; 95% CI 91.4-98.1%) and lower specificities (20.0%; 95% CI 11.2-33.0%). No significant difference
in performance was found when testing early versus late presenters; primary versus secondary cases; or DENV-1 versus DENV-2 infections. The use of a combination RDT fulfills WHO
ASSURED criteria for point-of-care testing and can enhance dengue diagnosis in an endemic setting. This has the potential to markedly improve clinical management of dengue in the field.
The Dengue IgM Capture ELISA (MAC-ELISA) is the immunoenzymatic system recommended by the Pan American Health Organization and the World Health Organization for the serological
diagnosis of dengue virus infection due to its high sensitivity; ease of performance; and use of a single acute-phase serum sample. However; tests with this enzyme-linked immunosorbent
assay (ELISA) system are time-consuming and require equipment for washing; incubation; and reading of the results. AuBioDOT is a multistep visual diagnostic immunoassay that uses
technology based on the immunoglobulin M (IgM) capture ELISA principle. This system uses white polyethylene opaque plates as the solid phase; colloidal gold as the marker; and silver ion
amplification. It does not require special equipment; it is totally manually operated; and it can be performed in less than 1 h. The sensitivity and specificity of AuBioDOT for the detection of
anti-dengue virus IgM antibodies were studied with a panel of 336 serum samples (150 serum samples from patients with suspected or serologically confirmed dengue virus infection; 186
serum samples from healthy blood donors and patients without dengue virus infection). The results were compared with those obtained by the MAC-ELISA. A sensitivity of 97.7% and a
specificity of 97.1% were obtained. The concordance of the two tests was 97.3%; with a kappa index of 0.94. The application of AuBioDOT for the detection of anti-dengue virus IgM
antibodies is recommended as an alternative method for the diagnosis of dengue virus infection; both for clinical diagnosis and for seroepidemiological surveillance. The system is useful
under field conditions and in laboratories and requires little equipment.
Background: Secondary dengue causes more severe disease than the primary. Early on; it is important to differentiate the two. We tried to find important clinical and laboratory differences
between the two for the purpose of early differentiation. Methods: One hundred fourteen patients confirmed on reverse transcriptase-polymerase chain reaction (RT PCR) were studied.
On day 2 of illness IgM and IgG indices were studied for calculation of IgG/IgM ratio. A one-step immunochromatographic assay was used for classification of patients into primary and
secondary dengue. Patient characteristics were also studied. Results: Dengue serotype 1 was the most common found in 60.5% patients. 66.7% (76 patients) had secondary dengue.
Secondary dengue cases had a higher mean temperature (101.56 ± 1.55 vs. 100.79 ± 1.25;°F; p 0.015); lower platelet counts (50.51 ± 38.91 vs. 100.45 ± 38.66; × 103/micl; p &lt,0.0001) and
a significantly higher percentage of Dengue hemorrhagic fever/Dengue shock syndrome (38.2% vs. 2.6%; p &lt,0.0001). In early phase of dengue NS1 and PCR were found to be better tests
for diagnosis and later IgM is better. The IgG/IgM ratio of ≥ 1.10 had a sensitivity of 100%; specificity of 97.4% and accuracy of 67.5% in differentiating secondary from primary dengue.
Conclusion: Early on in the clinical course; IgG/ IgM ratio can play an important role to differentiate the two. We found the ratio of ≥ 1.10 to be the best cut off for the same. © 2016 The
Author(s).
Background: Dengue fever is a frequent cause of admission to hospital in South East Asia; however many of the clinical characteristics and abnormalities on laboratory investigations at
presentation are found in other common infections. Objectives: To describe the clinical and laboratory features of dengue fever and other common febrile illnesses in Singapore. Study
design: We performed a prospective study of consecutive adult admissions to an infectious diseases hospital. Logistic regression analysis was used to identify symptoms; physical signs and
laboratory features that differentiated dengue fever from other febrile illnesses within the first 2 days of admission. Results: Of the 381 patients studied; 148 had serologically confirmed
dengue fever. Most of these had uncomplicated dengue fever; and only 9% had dengue haemorrhagic fever. A model based on clinical features alone (including a variety of cutaneous signs;
pulse rate and the presence of pharyngeal injection) was able to differentiate dengue fever from other infections with a sensitivity of 74% and specificity of 79%. A model based on clinical
features (rash) and laboratory parameters (white cell count; haemoglobin; prothrombin time; creatinine and bilirubin levels); achieved a sensitivity of 84% and specificity of 85%.
Conclusions: A combination of simple clinical and laboratory parameters are potentially able to predict dengue fever with a high level of accuracy in adults presenting to hospital with
febrile illnesses in Singapore. © 2005 Elsevier B.V. All rights reserved.
Dengue is known for its serious life-threatening complications. New rapid kits available recently in India target circulating non-structural protein (NS1) antigen from day one onwards. The
sensitivity and specificity of a newly introduced rapid combo kit against two conventional ELISA kits is assessed. The performance of this kit is quite satisfactory since excellent agreement of
94.26% was observed with particular reference to NS1 antigen detection among all three kits namely Rapid SD Bioline dengue Duo (SD Korea); InBios DENV Detect NS1 ELISA; USA and
dengue Early ELISA; Panbio; Australia. The false positivity of the rapid kit is very low since its specificity as for as NS1 antigen detection is concerned is 98.33%. The use of combination kit
helps to detect additional cases of dengue; which are negative for NS1 antigen but positive for IgM and/or IgG antibodies; thus facilitating early diagnosis in remote areas and small
laboratories.
A commercial Dengue Duo rapid test kit was evaluated for early dengue diagnosis by detection of dengue virus NS1 antigen and immunoglobulin M (IgM)/IgG antibodies. A total of 420
patient serum samples were subjected to real-time reverse transcription-polymerase chain reaction (RT-PCR); in-house IgM capture enzyme-linked immunosorbent assay (ELISA);
hemagglutination inhibition assay; and the SD Dengue Duo rapid test. Of the 320 dengue acute and convalescent sera; dengue infection was detected by either serology or RT-PCR in 300
samples (93.75%); as compared with 289 samples (90.31%) in the combined SD Duo NS1/IgM. The NS1 detection rate is inversely proportional; whereas the IgM detection rate is directly
proportional to the presence of IgG antibodies. The sensitivity and specificity in diagnosing acute dengue infection in the SD Duo NS1/IgM were 88.65% and 98.75%; respectively. The assay
is sensitive and highly specific. Detection of both NS1 and IgM by SD Duo gave comparable detection rate by either serology or RT-PCR.
High levels of anti-dengue IgM or IgG can be detected using numerous rapid diagnostic tests (RDTs). However; the sensitivity and specificity of these tests are reduced by changes in
envelope glycoprotein antigenicity that inevitably occur in limited expression systems. A novel RDT was designed to enhance diagnostic sensitivity. Dengue viruses cultured in animal cells
were used as antigens to retain the native viral coat protein. Monoclonal antibodies (mAbs) were then developed; for the first time; against domain I of envelope glycoprotein (EDI). The
anti-dengue EDI mAb was employed as a capturer; and EDII and EDIII; which are mainly involved in the induction of neutralizing antibodies in patients; were fully available to bind to anti-
dengue IgM or IgG in patients. A one-way automatic blood separation device prevented reverse migration of plasma and maximize the capture of anti-dengue antibodies at the test lines. A
clinical evaluation in the field proved that the novel RDT (sensitivities of 96.5% and 96.7% for anti-dengue IgM and IgG) is more effective in detecting anti-dengue antibodies than two major
commercial tests (sensitivities of 54.8% and 82% for SD BIOLINE, 50.4% and 75.3% for PanBio). The innovative format of RDT can be applied to other infectious viral diseases.
Introducción: el sistema comercial SD Dengue Duo (Standard Diagnostics) es una prueba rápida inmunocromatográfica que detecta la proteína NS1 del virus dengue y los anticuerpos anti-
dengue IgM e IgG de forma simultánea. Objetivo: evaluar las características funcionales y operacionales del sistema comercial para la detección de los diferentes marcadores virológicos y
serológicos. Métodos: se utilizó un panel constituido por 161 muestras de suero; 113 procedentes de pacientes con un diagnóstico clínico y serológico confirmado a cualquiera de los 4
serotipos del virus dengue y 48 negativas. Todas las muestras fueron procesadas por el sistema SD Dengue Duo y por las técnicas Platelia Dengue NS1 Ag; ELISA de captura IgM y el ELISA de
inhibición utilizadas como referencia. Resultados: el sistema evaluado mostró una sensibilidad de 57;75 por ciento para la proteína NS1; se observaron falsos negativos en muestras
colectadas a partir del quinto día de iniciados los síntomas en casos con infección secundaria. La detección de anticuerpos IgM mostró una sensibilidad de 96;0 por ciento y especificidad de

Evaluation of a capture
screening enzyme-linked
immunosorbent assay for
combined determination of
immunoglobulin M and G
antibodies produced during
dengue infection
Evaluation of a commercial
capture enzyme-linked
immunosorbent assay for
detection of immunoglobulin
M and G antibodies produced
during dengue infection
EVALUATION OF A
COMMERCIAL RAPID TEST KIT
FOR DETECTION OF ACUTE
DENGUE INFECTION
Evaluation of a commercially
available immunoglobulin M
capture enzyme- linked
immunosorbent assay kit for
diagnosing acute dengue
infections
Evaluation of a Dengue NS1
Antigen Detection Assay
Sensitivity and Specificity for
the Diagnosis of Acute Dengue
Virus Infection
Evaluation of a rapid
immunochromatographic
device for the detection of IgM
& IgG antibodies to Dengue
viruses (DENV) in a tertiary
care hospital in South India
Evaluation of a rapid
immunochromatographic test
in the diagnosis of dengue
fever
Evaluation of an
immunochromatography rapid
diagnosis kit for detection of
chikungunya virus antigen in
India; a dengue-endemic
98;4 por ciento. Se encontró una alta concordancia (95;7 por ciento) al clasificar el tipo de infección. En el estudio global de los 3 marcadores la sensibilidad del sistema se incrementó hasta
100 por ciento. Conclusiones: el SD Dengue Duo es un método simple; rápido; fácil de ejecutar; el cual no requiere de equipamiento adicional, tiene la ventaja de poder ser utilizado para
muestras; tanto de fase aguda como en la fase convaleciente y pudiera ser una alternativa para el diagnóstico del dengue en aquellos laboratorios que no cuenten con facilidades para ello.
A commercially available enzyme-linked immunosorbent assay (ELISA) (PanBio Dengue Screening ELISA) that utilized both immunoglobulin M (IgM) and IgG capture in the same microtiter
well for the diagnosis of dengue infection was evaluated. Sensitivity in primary and secondary dengue was 95%; while specificity was 94%.
A commercially available capture enzyme-linked immunosorbent assay (ELISA) for the detection of specific immunoglobulin M (IgM) and IgG antibodies produced during dengue infection
(PanBio Dengue Duo) was evaluated with paired serum specimens from 176 patients. Diagnosis was based on a hemagglutination inhibition (HAI) assay; with patients having either primary
dengue (n = 90); secondary dengue (n = 58); or no dengue (n = 28) infection. The combined use of IgM and IgG (sensitivity; 99%, specificity; 96%) was superior to the use of IgM alone
(sensitivity; 88%, specificity; 96%) or IgG alone (sensitivity; 85%, specificity; 96%). Furthermore; with the first serum sample of the pair of serum samples; the ELISA was able to diagnose
significantly more cases of dengue than the HAI assay (55% versus 14%). The results of the IgG capture ELISA gave a significant correlation with those of the HAI assay (r = 0.91, P < 0.0001);
and the IgG capture ELISA could be used to distinguish between primary and secondary infection. The best distinction was observed when an IgG cutoff ratio of 3.0 was used; with 88% of
primary infections and 98% of secondary infections being correctly classified. This ELISA should prove to be useful in the clinical diagnosis of dengue infection.
Early diagnosis is important for clinical management of dengue disease. While classic laboratory tests are often tedious and time consuming; point of care devices offer a rapid; cost-
effective and user-friendly alternative provided their accuracy is acceptable. This study evaluated the sensitivity; specificity and efficiency of SD BIOLINE Dengue Duo® rapid NS1; IgM and
IgG test kit for diagnosis of acute dengue virus infection. Standard laboratory diagnostics; RT-PCR; IgM and IgG capture ELISAs were carried out on 143 suspected dengue patient samples
obtained from a Sri Lankan population. Using the results of these standard laboratory tests as reference; the sensitivity and specificity of the SD Dengue Duo® NS1 test was 57% and 87%;
respectively; and those of the IgM test was 50% and 84%; respectively. The combined sensitivity and specificity of the SD Dengue Duo® NS1/ IgM test was 72% and 80%; respectively. The
SD Dengue Duo® NS1 test detected NS1 for up to 9 days from onset of fever. Primary and secondary dengue cases were classified according to the IgG test; of which the kit identified 88%
and 26% of primary and of secondary infection; respectively. Although the SD Dengue Duo® kit was not as accurate as the standard tests; it still can serve the useful reference for initial
screening of suspected dengue cases; especially in poor resource hospital settings and aid in clinical disease management of dengue infection.
Recently; commercially available kits for the detection of anti-dengue virus (anti-DEN) immunoglobulin M (IgM) antibodies have been developed. These standardized assays have greatly
enhanced our ability to effectively diagnose DEN infections. We conducted an evaluation of a test kit manufactured by MRL Diagnostics Inc. that is designed to detect anti-DEN IgM
antibodies. Eighty paired samples from DEN-infected individuals were tested by the MRL DEN Fever Virus IgM Capture enzyme-linked immunosorbent assay (ELISA); the PanBio Duo ELISA;
the PanBio Rapid Immunochromatographic Test (PRIT); and the IgM-IgG antibody capture (MAC/GAC) ELISA. All infections were confirmed by either PCR-assisted detection of DEN
transcripts or by DEN isolation in C6/36 cells. Seventeen paired samples from individuals with no evidence of acute DEN infection were used as negative controls. The PRIT had the best
sensitivity (100%); whereas the MAC/GAC ELISA and the PanBio Duo assay had the highest levels of specificity. The MRL ELISA and the PanBio Duo assay were the top performers when
taking into consideration both sensitivity and specificity. All assays were able to detect DEN-specific antibodies in samples from patients with either primary or secondary infections;
regardless of the infecting DEN serotype.
Background: Currently; no dengue NS1 detection kit has regulatory approval for the diagnosis of acute dengue fever. Here we report the sensitivity and specificity of the InBios DEN Detect
NS1 ELISA using a panel of well characterized human acute fever serum specimens. Methodology/Principal Findings: The InBios DENV Detect NS1 ELISA was tested using a panel composed
of 334 serum specimens collected from acute febrile patients seeking care in a Bangkok hospital in 2010 and 2011. Of these patients; 314 were found to have acute dengue by either RT-
PCR and/or anti-dengue IgM/IgG ELISA. Alongside the InBios NS1 ELISA kit; we compared the performance characteristics of the BioRad Platelia NS1 antigen kit. The InBios NS1 ELISA Ag kit
had a higher overall sensitivity (86% vs 72.8%) but equal specificity (100%) compared to the BioRad Platelia kit. The serological status of the patient significantly influenced the outcome. In
primary infections; the InBios NS1 kit demonstrated a higher sensitivity (98.8%) than in secondary infections (83.5%). We found significant variation in the sensitivity of the InBios NS1 ELISA
kit depending on the serotype of the dengue virus and also found decreasing sensitivity the longer after the onset of illness; showing 100% sensitivity early during illness; but dropping
below 50% by Day 7. Conclusion/Significance: The InBios NS1 ELISA kit demonstrated high accuracy when compared to the initial clinical diagnosis with greater than 85% agreement when
patients were clinically diagnosed with dengue illness. Results presented here suggest the accurate detection of circulating dengue NS1 by the InBios DENV Detect NS1 ELISA can provide
clinicians with a useful tool for diagnosis of early dengue infections. © 2014.
This study has evaluated the performance of a rapid immunochromatographic test (ICT) device in detecting antibodies to Dengue virus (DENV) in a tertiary hospital in South India. Sera from
hospital attendees; with requests for DENV antibody testing; were tested with the Panbio Dengue Duo Cassette and a reference antibody capture assay for the detection of IgM (Dengue
IgM capture ELISA-National Institute of Virology; India) and IgG (Dengue IgG capture ELISA-Panbio Diagnostics Inc.; Australia) antibodies. The ICT results were compared with results of
antibody capture tests for the detection of the IgM and IgG antibodies; respectively. Accuracy indices for IgM and IgG detection; respectively were- sensitivity 81.8% and 87.5%; specificity
75.0%; and 66.6%; positive predictive value (PPV) 61.0% and 72.9% and negative predictive value (NPV) 89.6% and 83.9%. The device performs poorly in detection of IgM and IgG antibodies
to DENVs and is not recommended for use as a stand-alone diagnostic test.
Serology is the mainstay of diagnosis in Dengue virus infection. Various rapid tests for antibody detection have been developed. They can prove to be important diagnostic tools especially
in the field set up due to technical simplicity. We evaluated a Rapid immunochromatographic assay. The rapid test was performed on acute phase sera collected from patients suspected to
be suffering from Denguefever/DHF. These samples were then tested by Dengue Duo Capture ELISA and compared. The rapid test showed a good sensitivity for the detection of secondary
dengue infection and thus can be a good screening tool.
Background: Chikungunya virus (CHIKV) and dengue virus (DENV) are arboviruses that share the same Aedes mosquito vector; and there is much overlap in endemic areas. In India; co-
infection with both viruses is often reported. Clinical manifestations of Chikungunya fever is often confused with dengue fever because clinical symptoms of both infections are similar. It is;
therefore; difficult to differentiate from those of other febrile illnesses; especially dengue fever. We previously developed a CHIKV antigen detection immunochromatography (IC) rapid
diagnosis kit [1]. The current study examined the efficacy of previously mentioned IC kit in India; a dengue-endemic country. Methods: Sera from 104 CHIKV-positive (by qRT-PCR) and/or
IgM-positive (ELISA) subjects collected in 2016; were examined. Fifteen samples from individuals with CHIKV-negative/DENV-positive and 4 samples from healthy individuals were also
 country
Evaluation of an
immunoglobulin M-specific
capture enzyme-linked
immunosorbent assay for
rapid diagnosis of dengue
infection
Evaluation of capture ELISA
and rapid
immunochromatographic test
for the determination of IgM
and IgG antibodies produced
during dengue infection
Evaluation of Commercially
Available Assays for Diagnosis
of Acute Dengue in
Schoolchildren during an
Epidemic Period in Medellin;
Colombia
Evaluation of Commercially
Available Diagnostic Tests for
the Detection of Dengue Virus
NS1 Antigen and Anti-Dengue
Virus IgM Antibody
Evaluation of immunoglobulin
M and G capture enzyme-
linked immunosorbent assay
Panbio kits for diagnostic
dengue infections
Evaluation of NS1Ag and IgM
antibodies against dengue;
importance for
epidemiological surveillance
Evaluation of OneStep Dengue
NS1 RapiDip™ InstaTest and
OneStep Dengue Fever
IgG/IgM RapiCard™ InstaTest
during the course of a dengue
type 1 epidemic
examined. Of the 104 CHIKV-positive sera; 20 were co-infected with DENV. Results: The sensitivity; specificity and overall agreement of the IC assay were 93.7; 95.5 and 94.3%; respectively;
using qRT-PCR as a gold standard. Also; there was a strong; statistically significant positive correlation between the IC kit device score and the CHIKV RNA copy number. The IC kit detected
CHIKV antigen even in DENV-co-infected patient sera and did not cross-react with DENV NS1-positive/CHIKV-negative samples. Conclusions: The results suggest that the IC kit is useful for
rapid diagnosis of CHIKV in endemic areas in which both CHIKV and DENV are circulating. © 2018 The Author(s).
Various assays have been developed to diagnose dengue virus infection; relying on techniques from the fields of serology and molecular biology. Many of these assays have been
successful; but there is still an urgent need for accurate; simple and rapid diagnostic assays to diagnose dengue virus infection and to assist in patient management. Using a panel of well-
characterized sera and a collection of retrospective samples obtained during the dengue epidemics that occurred in Belé;M Brazil; between 2002 and 2009; a modified immunoglobulin M-
specific capture enzyme-linked immunosorbent assay (Rapid-MAC-ELISA) was evaluated and compared with the "gold standard" MAC-ELISA; in order to assess the specificity; sensitivity;
stability; reproducibility and cost-effectiveness of this new assay. These results demonstrated that the Rapid-MAC-ELISA is comparable to the MAC-ELISA in terms of sensitivity and
specificity and is highly reproducible, additionally; it is easily performed; less expensive than other available formats and can be completed within three hours. Furthermore; the Rapid-
MAC-ELISA can be used for the diagnosis of dengue virus infections in resource-limited areas where dengue is endemic. © 2010 Elsevier B.V.
Background: Rapid diagnosis of dengue infection is essential to patient management and disease control. The development of a rapid (5 min) immunochromatographic test and a 2 h
commercial capture enzyme linked immunosorbent assay (ELISA) for anti-dengue IgM and IgG antibodies may lead to more rapid and accurate testing in peripheral health settings and
diagnostic laboratories. Objectives: Evaluate two new commercial tests for dengue serology (Dengue Rapid test and Dengue Duo ELISA, PanBio; Brisbane; Australia). Study design: The
sensitivity and specificity of the tests were compared with in-house dengue IgM ELISA and hemagglutination-inhibition (HAI) assays using known positive and negative dengue specimens;
as well as specimens from non-dengue cases. Results: Both assays showed excellent sensitivity in the diagnosis of both primary and secondary dengue infection (100%). In both assays; IgG
levels showed excellent correlation with hemagglutination-inhibition (HAI) assay; and these could be used to distinguish between primary and secondary dengue infections (92 and 97% of
patients correctly classified in the rapid test and Duo ELISA; respectively). Specificity in both assays was 89% when sera from patients; with no apparent dengue infection; typhoid;
leptospirosis and malaria; were tested. Conclusions: These tests should be a useful aid in confirming the clinical diagnosis of dengue infection. The rapid test will be particularly valuable in
peripheral health settings; while the ELISA has a place in central testing laboratories.
During an active surveillance study in school children in Medellin; we assessed the performance of two diagnostic strategies for dengue virus. A total of 41 patients with suspected dengue
acute infection were evaluated. Diagnostic strategies consisted of one combining Panbio (R) Dengue virus IgM and IgG Capture ELISAs (enzyme-linked immunosorbent assays) with reverse
transcriptase polymerase chain reaction (T-PCR) and another using a commercial rapid SD Bioline Dengue Duo (IgG/IgM + NS1 Ag) test. These two strategies were compared with the
enzyme linked immunospot microneutralization test (ELISPOT-MNT). The sensitivity and specificity were 53.9% and 80.0% for the combination of Panbio (R) ELISAs and RT-PCR tests; and
30.8% and 73.3% for the SD Bioline Duo test; respectively. ELISPOT-MNT detected 16.4% additional cases and revealed the presence of neutralizing antibodies in all the acute samples;
evidencing that they were all secondary infections. In contrast; Panbio (R) and SD Dengue Duo rapid tests only classified 23.0% and 26.9% of the cases as secondary dengue infections;
respectively. Cohen's kappa coefficient and McNemar's association tests demonstrated a significant disagreement between the two diagnostic strategies and ELISPOT-MNT. Overall; these
results evidence the relatively poor performances of commercial assays for the diagnosis of acute and secondary dengue infections; compared with ELISPOT-MNT; and raise concern about
the accuracy of these assays for the diagnostic of dengue in endemic areas.
Commercially available diagnostic test kits for detection of dengue virus (DENV) non-structural protein 1 (NS1) and anti-DENV IgM were evaluated for their sensitivity and specificity and
other performance characteristics by a diagnostic laboratory network developed by World Health Organization (WHO); the UNICEF/UNDP/World Bank/WHO Special Programme for
Research and Training in Tropical Diseases (TDR) and the Pediatric Dengue Vaccine Initiative (PDVI). Each network laboratory contributed characterized serum specimens for the panels
used in the evaluation. Microplate enzyme-linked immunosorbent assay (ELISA) and rapid diagnostic test (RDT formats) were represented by the kits. Each ELISA was evaluated by 2
laboratories and RDTs were evaluated by at least 3 laboratories. The reference tests for IgM anti-DENV were laboratory developed assays produced by the Armed Forces Research Institute
for Medical Science (AFRIMS) and the Centers for Disease Control and Prevention (CDC); and the NS1 reference test was reverse transcriptase polymerase chain reaction (RT-PCR). Results
were analyzed to determine sensitivity; specificity; inter-laboratory and inter-reader agreement; lot-to-lot variation and ease-of-use. NS1 ELISA sensitivity was 60–75% and specificity 71–
80%, NS1 RDT sensitivity was 38–71% and specificity 76–80%, the IgM anti-DENV RDTs sensitivity was 30–96%; with a specificity of 86–92%; and IgM anti-DENV ELISA sensitivity was 96–
98% and specificity 78–91%. NS1 tests were generally more sensitive in specimens from the acute phase of dengue and in primary DENV infection; whereas IgM anti-DENV tests were less
sensitive in secondary DENV infections. The reproducibility of the NS1 RDTs ranged from 92-99% and the IgM anti-DENV RDTs from 88–94%. © 2014.
Background: Serological assays are widely used to confirm dengue virus infections and to differentiate between a primary and a secondary infection. Objective: Two commercial dengue
diagnostic kits; Panbio Dengue IgM Capture and Dengue IgG Capture ELISA (Brisbane; Australia) were evaluated. Study design: Three hundred and seventy-three serum samples were
tested. Panel sera included samples from dengue confirmed cases (representing both primary and secondary infections); from non-dengue infectious diseases; and from healthy individuals.
The MAC-ELISA/Dengue IPK was used for the detection of anti-dengue virus IgM antibody in the sera and the ELISA inhibition method (EIM/Dengue IPK) was used to differentiate between
primary and secondary infections. Both these reference assays; which were previously developed in the Arbovirus Laboratory at the "Pedro Kouri" Tropical Medicine Institute; were
employed as the gold standard. Results: High sensitivity (96.8%) and specificity (99.4%) were found with the commercial diagnostics when compared to the reference methods.
Furthermore; high concordance 95.5% in classifying dengue infection types (primary or secondary infections) was observed. Conclusions: The Panbio Dengue IgM and IgG assays offer a
good alternative for dengue diagnosis. They are easy to perform and results can be obtained in less than 3 h. © 2007.
We determined the diagnostic performance of the OneStep NS1 and the OneStep IgG/IgM RDT kits against a panel of samples which comprised of 174 dengue positive and 165 dengue
negative sera characterized by three reference enzyme-linked immunosorbent assays (ELISAs). The diagnostic sensitivities of the OneStep kits for the detection of individual biomarkers of
NS1; IgM and IgG were 90% (95% CI: 82.1–94.7); 32.4% (95% CI: 24.8–40.8) and 44.4% (95% CI: 38.2–50.7); respectively. The combination of the OneStep IgG/IgM kit with the OneStep NS1
kit demonstrated significantly higher sensitivities for the combined NS1/IgM (96.8%, 95% CI: 90.9–99.3) and NS1/IgM/IgG (99.5%, 95% CI: 97.1–99.9)(P < 0.001). In conclusion; the OneStep
NS1 kit has high sensitivity and specificity and is highly recommended for use. The low sensitivities for IgG (44.4%) and for IgM (32.4%) of the OneStep IgG/IgM kit when used alone suggest
it is best used in combination with the OneStep NS1 kit to enhance its overall diagnostic performance. © 2017 Elsevier Inc.

Evaluation of six commercial
point-of-care tests for
diagnosis of acute dengue
infections: The need for
combining NS1 antigen and
IgM/IgG antibody detection to
achieve acceptable levels of
accuracy
Evaluation of six
immunoassays for detection of
dengue virus-specific
immunoglobulin M and G
antibodies
Evaluation of the
performances of six
commercial kits designed for
dengue NS1 and anti-dengue
Ig;M IgG and IgA detection in
urine and saliva clinical
specimens
Evaluation of the SD BIOLINE
dengue duo rapid test in the
course of acute and
convalescent dengue
infections in a Mexican
endemic region
Evaluation of the SD Dengue
Duo diagnosis system for
detection of NS1 protein and
IgM and IgG dengue
antibodies
Expression on dengue virus
type 2 envelope glycoprotein
and application of
recombinant protein in
serologic diagnosis.
Six assays were evaluated in this study to determine their suitability for the diagnosis of acute dengue infection using samples from 259 Sri Lankan patients with acute fevers (99 confirmed
dengue cases and 160 patients with other confirmed acute febrile illnesses): (i) the Merlin dengue fever IgG & IgM combo device (Merlin); (ii) the Standard Diagnostics Dengue Duo
nonstructural 1 (NS1) antigen and IgG/IgM combo device (Standard Diagnostics; South Korea); (iii) the Biosynex Immunoquick dengue fever IgG and IgM (Biosynex; France) assay; (iv) the
Bio-Rad NS1 antigen strip (Bio-Rad; France); (v) the Panbio Dengue Duo IgG/IgM Cassette (Inverness; Australia); and (vi) the Panbio dengue NS1 antigen strip (Inverness; Australia). The
median number of days of fever prior to admission sample collection was 5 days (interquartile range; 3 to 7 days). Sensitivity and specificity of the NS1 antigen tests ranged from 49 to 59%
and from 93 to 99%; respectively; and sensitivity and sensitivity of the IgM antibody test ranged from 71 to 80% and from 46 to 90%; respectively. Combining the NS1 antigen and IgM
antibody results from the Standard Diagnostics Dengue Duo test gave the best compromise of sensitivity and specificity (93% and 89%; respectively) and provided the best sensitivity in
patients presenting at different times after fever onset. The Merlin IgM/IgG antibody tests correctly classified 64% and 86% of the primary and secondary dengue infection cases;
respectively; and the Standard Diagnostics IgM/IgG antibody tests correctly classified 71% and 83% of the primary and secondary dengue infection cases; respectively. This study provides
strong evidence of the value of combining dengue antigen- and antibody-based test results in the rapid diagnostic test (RDT) format for the acute diagnosis of dengue. Copyright © 2011;
American Society for Microbiology. All Rights Reserved.
The performance of six commercially available immunoassay systems for the detection of dengue virus-specific immunoglobulin M (IgM) and IgG antibodies in serum was evaluated. These
included two IgM and IgG enzyme immunoassays (EIA) from MRL Laboratories and PanBio; a rapid immunochromatographic test (RIT) from PanBio; immunofluorescence assays (IFA) from
Progen; a dot blot assay from Genelabs; and a dipstick EIA from Integrated Diagnostics (INDX). For this study a panel of 132 serum samples; including 90 serum samples from patients with
suspected dengue virus infection and 42 serum samples from patients with other viral infections; was used. In addition; serial serum samples from two monkeys experimentally immunized
and challenged with dengue virus type 2 were used. Results were considered conclusive when concordant results were obtained with four of the six antibody-specific assays. Based on this
definition; the calculated overall agreement for the human serum samples for the respective IgM immunoassays was 97% (128 of 132); with 34% (45 of 132) positive serum samples; 63%
(83 of 132) negative samples; and 3% of samples (4 of 132) showing discordant results. The calculated overall agreement for the IgG assays was 94% (124 of 132); with 49% (65 of 132)
positive; 45% (59 of 132) negative; and 6% (8 of 132) discordant results; respectively. The sensitivities of the dengue virus-specific assays evaluated varied between 71 and 100% for IgM
and between 52 and 100% for IgG; with specificities of 86 to 96% and 81 to 100%; respectively. The relative sensitivities of the respective IgM assays measured with the monkey serum
samples were comparable with those obtained with 12 serial serum samples from humans. Overall performance; based on the sum of the agreement; sensitivity; specificity; and Kappa
statistics of the IgM and IgG immunoassays; showed that the antibody detection systems from INDX and Genelabs and the MRL and PanBio EIA are useful and reliable assays for dengue
virus serodiagnosis.
Background: Rapid diagnostic tests (RDTs) have been commercialized in order to help physicians in dengue diagnosis. Until recently; only blood samples were used for those tests but it has
been shown in several studies that urine and saliva can also be employed for dengue diagnosis. RDTs for the detection of NS1 antigen and anti-dengue IgG; IgM and IgA in urine and saliva
specimens have thus been developed by Standard Diagnostics. The aim of this study was to evaluate the performances these new commercial assays. Methods: Two panels of clinical
specimens were used: one for the evaluation of the NS1-detection devices and the second for the evaluation of the antibody-detection kits. Each panel consisted of urine and saliva
specimens collected sequentially from 86 patients with a confirmed dengue infection. A total of 291 saliva and 440 urine samples were included in the NS1-evaluation panel and 530 saliva
and 528 urine specimens constituted the antibody-evaluation panel. All samples were tested in parallel by in-house ELISAs and by the commercial RDTs. Results: The RDTs demonstrated an
overall sensitivity of 15.5 %/27.9 %/10.7 % for NS1/IgG/IgA detection in urine samples and 20.4 %/ 34.8 %/11 %/6.2 % for NS1/IgG/IgM/IgA detection in saliva samples. Compared to the in-
house NS1 ELISA; the results obtained with the NS1 RDT demonstrated a good correlation with urine samples (kappa coefficient: 0.88) but not with saliva specimens (kappa coefficient:
0.28). RDTs designed for antibody detection in saliva and urine were extremely specific (100 %); but less sensitive than the in-house ELISAs (i.e.; reduction of the overall sensitivity by 12.2 %
for the RDT designed for IgG detection in urine and by 23.7 % for the RDT detecting anti-DENV IgM in saliva). IgM were not detected in urine; either by RDT or ELISA. Conclusions: Although
the RDTs evaluated here offer an apparently attractive approach for dengue diagnosis; this study suggests that these new commercial kits would require further improvement to increase
the sensitivity. © 2016 Andries et al.
In this study; we evaluated the performance of a rapid test; the SD BIOLINE Dengue Duo (SD BDD) kit; with a panel of serum samples from 310 Mexican patients with diagnosis of dengue
infection previously confirmed by reference enzyme-linked immunosorbent assay tests. Eighty-seven negative samples from other febrile illnesses were included as controls. The SD BDD
showed an overall sensitivity of 90.65% and specificity of 89.66%. No statistically significant differences were found in the sensitivity of the SD BDD kit compared between primary or
secondary infections (87.05% versus 93.57%; respectively; P = 0.0761) and dengue fever or dengue hemorrhagic fever cases (90.77% versus 89.74%; respectively; P = 0.7716). However; a
higher sensitivity in the acute phase of dengue infection was found compared with the convalescent phase (93.03% versus 81.82%; respectively; P = 0.0089). These results indicate that the
SD BDD kit is a useful tool to diagnose dengue infections; both in primary or secondary infections and mainly during the acute phase. (C) 2014 Elsevier Inc. All rights reserved.
Introduction: SD Dengue Duo (Standard Diagnosis) commercial kit is an immunochromatographic rapid test that detects NS1 protein and IgG/IgM dengue antibodies simultaneously.
Objective: to evaluate the operational and functional characteristics of this system for the detection of virological and serological markers. Methods: sera panel was made up by 161
samples; 113 from patients with clinically and serologically confirmed dengue caused by any of the four dengue virus serotypes and 48 negative samples. All these samples were tested by
SD Dengue Duo Kit and by Platelia Dengue NS1 Ag; IgM Capture ELISA and ELISA Inhibition Method used as reference assays. Results: the evaluated kit showed a 57.75% sensitivity for the
detection of NS1 protein; false negatives were detected in samples collected 5 days or more after fever onset in secondary infection cases. IgM detection showed 96.0% sensitivity and
98.4% specificity. Furthermore; high agreement (95.7%) in classifying dengue infection types (primary or secondary infections) was observed. The global study of the 3 markers; the
sensitivity rose to 100%. Conclusions: SD Dengue Duo is a simple; easy and rapid assay, it does not require additional equipment; can be used for acute and convalescence serum samples
and offers a good alternative for dengue diagnosis in those laboratories where a complete dengue virus diagnosis is difficult to perform.
To obtain dengue virus type 2 envelope glycoprotein for serological diagnosis of dengue virus infection; the truncated E protein deleting the carboxy-terminal -100 aa of dengue virus type 2
was amplified by reverse transcription polymerase chain reaction (PCR). The products of PCR were ligated with plasmid pET-30a and transformed into Escherichia coli competent cells BL21
(DE3) to express recombinant protein. The expressed product was purified by electroelution. The purified recombinant protein was applied to test sera from patients infected with dengue
virus type 2 by indirect enzyme linked immunosorbent assay (ELISA). The results were compared with those tested by IFA or Panbio Dengue IgG capture ELISA. Results showed that the
recombinant plasmid was successfully constructed and expressed in E. coli. After the recombinant protein was used to detect sera from patients infected with dengue virus type 2; a
sensitivity of 95.6% and an accuracy of 96.8% were obtained by comparing with IFA. Furthermore; the results were in excellent agreement with those obtained by using a commercially
available diagnostic kit; Dengue Duo rapid strip test from Panbio. In conclusion; the truncated E protein dengue virus type 2 was successfully expressed in E. coli and the recombinant
protein could be applied as antigens for dengue serum detection.
Facilitation of Cell Adhesion by
Immobilized Dengue Viral
Nonstructural Protein 1 (NS1):
Arginine-Glycine-Aspartic Acid
Structural Mimicry within the
Dengue Viral NS1 Antigen.
Field Evaluation and Impact on
Clinical Management of a
Rapid Diagnostic Kit That
Detects Dengue NS1; IgM and
IgG
Immunoglobulin G (IgG) to IgM
ratio in secondary adult
dengue infection using
samples from early days of
symptoms onset
International external quality
control assessment for the
serological diagnosis of
dengue infections
Interpretation of the presence
of IgM and IgG antibodies in a
rapid test for dengue: Analysis
of dengue antibody prevalence
in Fortaleza City in the 20th
year of the epidemic
Introduction of a rapid
diagnostic dengue NS1
antigen–IgM/IgG combination
assay associated with a
reduced inpatient length of
stay
Laboratory diagnosis of
dengue: A review
: Dengue virus infection causes life-threatening hemorrhagic fever. Increasing evidence implies that dengue viral nonstructural protein 1 (NS1) exhibits a tendency to elicit potentially
hazardous autoantibodies; which show a wide spectrum of specificity against extracellular matrix and platelet antigens. How NS1 elicits autoantibodies remains unclear. To address the
hypothesis that NS1 and matrix proteins may have structural and functional similarity; cell-matrix and cell-NS1 interactions were evaluated using a cell-adhesion assay. The present study
showed that dengue NS1 immobilized on coverslips resulted in more cell adhesion than did the control proteins. This cell adhesion was inhibited by peptides containing arginine-glycine-
aspartic acid (RGD); a motif important for integrin-mediated cell adhesion. In addition; anti-NS1 antibodies blocked RGD-mediated cell adhesion. Although there is no RGD motif in the NS1
protein sequence; these data indicate that RGD structural mimicry exists within the NS1 antigen; (C) Copyright Oxford University Press 2002.
Background: Dengue diagnosis is complex and until recently only specialized laboratories were able to definitively confirm dengue infection. Rapid tests are now available commercially
making biological diagnosis possible in the field. The aim of this study was to evaluate a combined dengue rapid test for the detection of NS1 and IgM/IgG antibodies. The evaluation was
made prospectively in the field conditions and included the study of the impact of its use as a point-of-care test for case management as well as retrospectively against a panel of well-
characterized samples in a reference laboratory. Methodology/Principal Findings: During the prospective study; 157 patients hospitalized for a suspicion of dengue were enrolled. In the
hospital laboratories; the overall sensitivity; specificity; PPV and NPV of the NS1/IgM/IgG combination tests were 85.7%; 83.9%; 95.6% and 59.1% respectively; whereas they were 94;4%;
90.0%; 97.5% and 77.1% respectively in the national reference laboratory at Institut Pasteur in Cambodia. These results demonstrate that optimal performances require adequate training
and quality assurance. The retrospective study showed that the sensitivity of the combined kit did not vary significantly between the serotypes and was not affected by the immune status
or by the interval of time between onset of fever and sample collection. The analysis of the medical records indicates that the physicians did not take into consideration the results obtained
with the rapid test including for care management and use of antibiotic therapy. Conclusions: In the context of our prospective field study; we demonstrated that if the SD Bioline Dengue
Duo kit is correctly used; a positive result highly suggests a dengue case but a negative result doesn't rule out a dengue infection. Nevertheless; Cambodian pediatricians in their daily
practice relied on their clinical diagnosis and thus the false negative results obtained did not directly impact on the clinical management. © 2012 Andries et al.
Background: Dengue virus (DENV) infection is an emerging arboviral infection in tropical and sub-tropical countries in South-East Asia; the Western Pacific and South and Central America.
Secondary DENV infection is the most widely accepted risk factor for the development of severe forms. Methods to discriminate primary and secondary DENV infection may be of great
prognostic value. ELISA based detection of specific antibodies (IgG and IgM) to the four DENV serotypes is valuable for detemination of primary or secondary infection. Several studies had
been performed to discriminate primary and secondary DENV infection using the ratio of IgG and IgM at the various days of symptoms onset. The aim of this study is to determine the best
cut-off point of IgG to IgM ratio is able to discriminating secondary from primary DENV infection in adult using samples from early days of symptoms onset. Methods: This prospective
cohort study on 48 adult patients with DENV infected patients on the period of August 2011-January 2012 in 5 out-patient settings health facilities in Tangerang district; Banten province;
Indonesia with chief complaint of fever less than 3 days. Datas were collected on the day the patients attended health facilities; consisted of demographic; clinical; laboratory; and
serological data. Serological data from 48 serum sample from adult patients were evaluated using Focus Diagnostics Dengue Virus IgM and IgG Capture DxSelect (TM) ELISA Kits to
determine IgG; IgM index values and SD Bioline Dengue Duo (TM) Rapid Tests to determine NS1; IgG; and IgM result. Results: According to NS1; IgG and IgM results; 36 patients were
classified as secondary infection; 12 were primary Infection. The mean (SD) of IgG/IgM ratios for secondary and primary infection were 3.28 (0.54) and 0.18 (0.11) consecutively. The
IgG/IgM ratio of >= 1.14 confirmed secondary infection with sensitivity of 80.56 %; specificity 91.67 %; accuracy level 83.33 %; and likely hood ratio of (LR) 9.67. Conclusion: The IgG/IgM
ratio of >= 1.14 as the best cut off point to determine secondary DENV infection in early days of symptoms onset.
Background: Dengue is endemic to the tropics and subtropics; and the most frequent of arthropod-borne viral diseases. Reliable diagnosis of dengue infection is important not only in
clinical care but also in disease surveillance; the control of outbreaks; and the development of new vaccines. The diagnosis of dengue infection is usually established by a variety of
commercial or in-house serological protocols. The European Network for the Diagnostics of Imported Viral Diseases (ENIVD) recognized the need to survey the accuracy of dengue
serological diagnostics in current use; and organized an external quality assurance (EQA) study of dengue serological practice in diagnostic laboratories. Methods: A 15-sample panel;
consisting of sera reactive against dengue plus specificity and negative controls; was sent to 48 laboratories for serological testing. The results returned by the participating laboratories
were anonymized; scored; and subjected to comparison and statistical analysis. Results: Ten laboratories rated all samples correctly with regard to Ig;M and only three achieved the full
score for IgG detection. The main handicaps in assay performance were suboptimal sensitivity of in-house IgM detection protocols by comparison with better-performing commercial ELISA
tests; and the presence of IgG cross-reactivity with heterologous flaviviruses. Differences of detail in the methodology of dengue IgG antibody detection appear to underlie the disparities in
accuracy observed between laboratories. Conclusion: This EQA study demonstrates that there is room for many laboratories to improve sensitivity in the detection of anti-dengue virus IgM
antibodies; against the benchmark set by commercial antibody capture ELISA tests. The EQA shows also that cross-reactivity is a continuing issue; and IgG detection protocols must be
optimized to increase their specificity. © 2015 Domingo et al.
Introduction: The diagnosis of dengue and the differentiation between primary and secondary infections are important for monitoring the spread of the epidemic and identifying the risk of
severe forms of the disease. The detection of immunoglobulin (Ig)M and IgG antibodies is the main technique for the laboratory diagnosis of dengue. The present study assessed the
application of a rapid test for dengue concerning detection of new cases; reinfection recognition; and estimation of the epidemic attack rate. Methods: This was a retrospective; cross-
sectional; descriptive study on dengue using the Fortaleza Health Municipal Department database. The results from 1;530 tested samples; from 2005-2006; were compared with data from
epidemiological studies of dengue outbreaks in 1996; 2003; and 2010. Results: The rapid test confirmed 52% recent infections in the tested patients with clinical suspicion of dengue: 40%
detected using IgM and 12% of new cases using IgG in the non-reactive IgM results. The positive IgM plus negative IgG (IgM+ plus IgG-) results showed that 38% of those patients had a
recent primary dengue infection; while the positive IgG plus either positive or negative IgM (IgG+ plus IgM+/-) results indicated that 62% had dengue for at least a second time (recent
secondary infections). This proportion of reinfections permitted us to estimate the attack rate as ≥62% of the population sample. Conclusions: The rapid test for dengue has enhanced our
ability to detect new infections and to characterize them into primary and secondary infections; permitting the estimation of the minimal attack rate for a population during an outbreak.
Dengue is an arthropod borne disease that has become important worldwide. There is still no specific drug available for treatment and also no protective vaccine that can be used. As such;
specific diagnosis is essential to enable good management and prevention of large outbreaks. Diagnosis today in many countries is still based on serology though the detection of NS1 has
slowly become incorporated. Diagnosis is critical for early intervention with specific preventive health measures to prevent fatalities and also to curtail spread and reduce economic losses.

Low specificity of an
immunochromatographic
serological assay for diagnosis
of dengue fever in travelers
returning with malaria.
Modeling of hemoglobin in
dengue fever and dengue
hemorrhagic fever using
bioelectrical impedance
Multicountry Prospective
Clinical Evaluation of Two
Enzyme-Linked
Immunosorbent Assays and
Two Rapid Diagnostic Tests for
Diagnosing Dengue Fever
New dengue antibody assay
with unique differential
detection of IgG and IgM
antibodies
NS1-based tests with
diagnostic utility for
confirming dengue infection: a
meta-analysis
Performance of Simplexa
Dengue molecular assay
compared to conventional and
SYBR Green RT-PCR for
detection of dengue infection
in Indonesia
Serological assays mainly detect IgM which now as a single test is invalid unless a second sample is taken to confirm. As such to effectively diagnose dengue at all stages of infection; assays
with two or more markers are required or two samples taken a few days apart. Other commonly used tests include NS1 detection; nucleic acid amplification and IgG detection. However the
sensitivities of the current commercial kits vary quite considerably and have to be interpreted with caution. Hence knowledge of this disease is essential when conducting diagnostics for
dengue.
The dengue duo rapid strip test (DDRST) was evaluated with 37 travellers admitted for febrile syndrome at the University Hospitals of Marseilles; France; with blood smear positive for
Plasmodium falciparum. The patients were hospitalized between October 2001 and April 2002 after returning from the Comoros islands (n=21); Madagascar (n=1); West Africa (n=9); East
Africa (n=1); India (n=1); French Polynesia (n=2) and West Indies (n=2). Eighteen sera tested positive for the presence of immunoglobulin (Ig)M. Of these; only two were confirmed by ELISA
(54.3% specificity). All 19 sera testing negative for IgM with DDRST were found negative with ELISA (100% negative predictive value). Positive predictive value was 11.1%. The patients who
exhibited a false-positive IgM detection by DDRST were returning from Comoros Islands (n=13); East Africa (n=2) and West Indies (n=1). It is concluded that the elevated rate of false-
positive IgM detected by DDRST is certainly not an immunological cross-reactivity due to poor specificity of the recombinant envelope protein.
This paper describes a model for predicting hemoglobin (Hb) by using bioelectrical impedance analysis (BIA) in dengue patients in the Hospital Universiti Kebangsaan Malaysia (HUKM).
Bioelectrical impedance measurements were conducted on 83 (47 males and 36 females) serologically confirmed dengue fever (DF) and dengue hemorrhagic fever (DHF) patients during
their hospitalization. The predictive equation for Hb was derived using multivariate analysis. We investigated all the parameters in BIA; patients' symptom and demographic data. In this
developed model; four predictors (reactance (Xc); sex; weight and vomiting) were found to be the best predictive factors for modeling Hb in dengue patients. However; the model can only
explain approximately 42% of the variation in Hb status; thus single frequency bio-impedance stand-alone technique is insufficient to monitor Hb for the DF and DHF patients. Further
investigation using multi-frequency BIA is recommended in modeling Hb to achieve the most parsimonious model.
We evaluated four dengue diagnostic devices from Alere; including the SD Bioline Dengue Duo (nonstructural [NS] 1 Ag and IgG/IgM); the Panbio Dengue Duo Cassette (IgM/IgG) rapid
diagnostic tests (RDTs); and the Panbio dengue IgM and IgG capture enzyme-linked immunosorbent assays (ELISAs) in a prospective; controlled; multicenter study in Peru; Venezuela;
Cambodia; and the United States; using samples from 1;021 febrile individuals. Archived; well-characterized samples from an additional 135 febrile individuals from Thailand were also
used. Reference testing was performed on all samples using an algorithm involving virus isolation; in-house IgM and IgG capture ELISAs; and plaque reduction neutralization tests (PRNT) to
determine the infection status of the individual. The primary endpoints were the clinical sensitivities and specificities of these devices. The SD Bioline Dengue Duo had an overall sensitivity
of 87.3% (95% confidence interval [CI]; 84.1 to 90.2%) and specificity of 86.8% (95% CI; 83.9 to 89.3%) during the first 14 days post-symptom onset (p.s.o.). The Panbio Dengue Duo Cassette
demonstrated a sensitivity of 92.1% (87.8 to 95.2%) and specificity of 62.2% (54.5 to 69.5%) during days 4 to 14 p.s.o. The Panbio IgM capture ELISA had a sensitivity of 87.6% (82.7 to
91.4%) and specificity of 88.1% (82.2 to 92.6%) during days 4 to 14 p.s.o. Finally; the Panbio IgG capture ELISA had a sensitivity of 69.6% (62.1 to 76.4%) and a specificity of 88.4% (82.6 to
92.8%) during days 4 to 14 p.s.o. for identification of secondary dengue infections. This multicountry prospective study resulted in reliable real-world performance data that will facilitate
data-driven laboratory test choices for managing patient care during dengue outbreaks.
Objectives: Most of the serological methods; traditional and recent; that have been developed to assess laboratory confirmation of dengue infection; suffer from the need to use separate
tests for IgM and for IgG and the need for paired serum specimens in order to establish a clinical interpretation. Moreover; most of the IgG tests ignore early convalescence as well as past
exposure. The objectives of this study is to demonstrate the advantages of the ImmunoComb Dengue BiSpot; a new dengue antibody assay; that detects simultaneously both dengue-
specific IgM and IgG; in determining type and status of infection in a single test. Designs and methods: Single and paired positive and negative specimens obtained from various dengue-
high prevalent regions and from dengue-free areas were used. The specimens were comprised of admission samples of suspected patients and secondary dengue infection individuals; a
group of hospital patients; asymptomatic to dengue who originated from dengue high prevalence areas and dengue-negative volunteers. All samples were tested by the ImmunoComb
Dengue BiSpot kit along with the gold standard assays of Hemagglutination-inhibition (HAI) and MAC ELISA. Results: Testing 365 gold standard positive specimens from different regions;
the ImmunoComb Dengue BiSpot kit revealed sensitivity for IgM of 97.5% and 99.1% for IgG. The IgG was segregated into 2 groups: 86.6% for high titer and 12.5% for low titer of IgG. The
overall sensitivity was 98.9%. The overall specificity of the kit; testing 657 samples; mostly from dengue-free area was 97.9%. The higher diagnostic resolution of the Dengue BiSpot test was
demonstrated when the performance of the kit on the various groups was compared to the gold standard tests and the dengue status was determined in paired specimens by the new
assay. Conclusions: The results of the present studies demonstrate the advantages of ImmunoComb Dengue BiSpot assay as an alternate strategy for the determination of dengue infection
status. The combined use of IgM and IgG to discriminate between primary and secondary infections; on one hand; and providing high resolution in distinction of convalescence stage and
past infection; on the other hand; endow the kit with improved capacity in assessing the clinical status of a tested individual in a single test. © 2008 The Canadian Society of Clinical
Chemists.
Objectives: Non-structural protein 1 (NS1)-based tests may offer a larger window of opportunity for dengue diagnosis and could constitute a very useful diagnostic tool. The aim of this
study was to establish the overall accuracy of NS1-based tests for diagnosing dengue infection. Methods: A meta-analysis was conducted including 18 studies published up to October 1;
2012 identified using PubMed; ISI Web of Science; Google Scholar; and the Chinese National Knowledge Infrastructure (CNKI) database. Results: For the single NS1-based tests - ELISA
(Panbio Dengue Early ELISA Kit; Dengue NS1 Ag ELISA Kit; and Platelia Dengue NS1 Ag-ELISA Kit) and immunochromatography (Dengue NS1 Ag STRIP Kit and SD BIOLINE Dengue Duo Strip
Kit) - the summarized sensitivities and specificities were 67% (95% confidence interval (CI) 59-74%) and 99% (95% CI 97-99%); and 71% (95% CI 61-79%) and 99% (95% CI 98-100%);
respectively. The hierarchical summary receiver operating characteristics (HSROCs) were 0.92 and 0.96; respectively. For NS1 combined with an anti-dengue-specific IgM test; the
summarized sensitivity; specificity; and HSROC were 83% (95% CI 68-92%); 86% (95% CI 79-91%); and 0.91 (95% CI 0.89-0.93); respectively. The accuracy for serotypes was 50.0-90.9% for
DENV-1; 38.5-85.7% for DENV-2; 46.7-91.3% for DENV-3; and 21.7-87.0% for DENV-4. Conclusions: These results support the use of single NS1-based tests, they have good diagnostic utility
for confirming dengue and for distinguishing serotypes DENV-1 and 3 from DENV-2 and 4; while they can be used as a screening tool when combined with an IgM test. Moreover; the
Dengue NS1 Ag STRIP Kit appears to be the best for confirming and serotyping dengue infection. (C) 2014 The Authors. Published by Elsevier Ltd on behalf of International Society for
Infectious Diseases. This is an open access article under the CC BY-NC-ND license.
Diagnostic tests based on detection of dengue virus (DENV) genome are available with varying sensitivities and specificities. The Simplexa Dengue assay (Focus Diagnostics) is a newly
developed real-time RT-PCR method designed to detect and serotype DENV simultaneously. To assess the performance of the Simplexa Dengue assay; we performed comparison with
conventional RT-PCR and SYBR Green real-time RT-PCR on patients sera isolated from eight cities across Indonesia; a dengue endemic country. A total of 184 sera that were confirmed using
NS1 and/or IgM and IgG ELISA were examined. Using conventional and SYBR Green real-time RT-PCR; we detected DENV in 53 (28.8%) and 81 (44.0%) out of 184 sera; respectively. When
the Simplexa Dengue assay was employed; the detection rate was increased to 76.6% (141 out of 184 samples). When tested in 40 sera that were confirmed by virus isolation as the gold
standard; the conventional RT-PCR yielded 95% sensitivity while the sensitivity of SYBR Green real-time RT-PCR and Simplexa Dengue assay reached 97.5% and 100%; respectively. The
specificities of all methods were 100% when tested in 43 non-dengue illness and 20 healthy human samples. Altogether; our data showed the higher detection rate of Simplexa Dengue

Prospective evaluation of the
SD BIOLINE Dengue Duo rapid
test during a dengue virus
epidemic
Quantification of NS1 dengue
biomarker in serum via
optomagnetic nanocluster
detection
Rapid antigen tests for dengue
virus serotypes and zika virus
in patient serum
Rapid diagnosis of dengue
outbreaks in resource limited
facilities
Rapid diagnostic tests for
dengue and leptospirosis:
antibody detection is
insensitive at presentation.
Rapid Diagnostic Tests for
Dengue Virus Infection in
Febrile Cambodian Children:
Diagnostic Accuracy and
Incorporation into Diagnostic
Algorithms
Rapid diagnostic tests for non-
malarial febrile illness in the
tropics
compared to conventional and SYBR Green real-time RT-PCR in field/surveillance setting. In conclusion; Simplexa Dengue offers rapid and accurate detection and typing of dengue infection
and is suitable for both routine diagnostic and surveillance. © 2014 Sasmono et al.
Dengue virus is endemic in French Guiana with occurrence of cyclical outbreaks. There is a need for rapid tests allowing dengue laboratory diagnosis in healthcare centers scattered
throughout this wide Amazonian territory. Our objective was to evaluate the real-life performance of the SD BIOLINE Dengue Duo (IgG/IgM + NS1 Ag) rapid test (RDT) during the 2012–2013
dengue epidemics. The RDT was evaluated in parallel with routine laboratory tests; PlateliaTM Dengue NS1 Ag and Focus Diagnostics Dengue Fever Virus IgM Capture DxSelect. A total of
3;347 patients with suspected dengue acute infection were evaluated. The diagnostic performances of the SD BIOLINE NS1 Ag were equivalent to Platelia NS1; 471 patients (14.1%) were
NS1 Ag positive with the RDT and 14.2% with Platelia. The Cohen’s Kappa coefficient was 0.86 *95%CI: 0.83–0.88]; indicating an almost perfect agreement. Moreover; the sensitivity of SD
BIOLINE NS1 Ag relative to the RT-PCR method was 87% [95%CI: 80–93%] and the specificity was 92% [95% CI: 87–97%]. However; the SD BIOLINE IgM test was found positive in 6.3% of the
samples in comparison to 10.7% with Dx Select IgM. The Cohen’s Kappa coefficient was 0.53 *95%CI: 0.47–0.58] indicating a moderate agreement. This raised concern about the SD BIOLINE
IgM for the diagnostic of dengue in endemic areas. When considering only NS1 Ag results and not Ig;M the RDT could be a viable solution to manage dengue outbreaks in healthcare
centers where no laboratory services are available; in the early phase of the disease. © 2017; Springer-Verlag GmbH Germany.
Dengue is a tropical vector-borne disease without cure or vaccine that progressively spreads into regions with temperate climates. Diagnostic tools amenable to resource-limited settings
would be highly valuable for epidemiologic control and containment during outbreaks. Here; we present a novel low-cost automated biosensing platform for detection of dengue fever
biomarker NS1 and demonstrate it on NS1 spiked in human serum. Magnetic nanoparticles (MNPs) are coated with high-affinity monoclonal antibodies against NS1 via bio-orthogonal Cu-
free 'click' chemistry on an anti-fouling surface molecular architecture. The presence of the target antigen NS1 triggers MNP agglutination and the formation of nanoclusters with rapid
kinetics enhanced by external magnetic actuation. The amount and size of the nanoclusters correlate with the target concentration and can be quantified using an optomagnetic readout
method. The resulting automated dengue fever assay takes just 8 minutes; requires 6 μL of serum sample and shows a limit of detection of 25 ng/mL with an upper detection range of
20000 ng/mL. The technology holds a great potential to be applied to NS1 detection in patient samples. As the assay is implemented on a low-cost microfluidic disc the platform is suited for
further expansion to multiplexed detection of a wide panel of biomarkers.
The recent Zika virus (ZIKV) outbreak demonstrates that cost-effective clinical diagnostics are urgently needed to detect and distinguish viral infections to improve patient care. Unlike
dengue virus (DENV); ZIKV infections during pregnancy correlate with severe birth defects; including microcephaly and neurological disorders. Because ZIKV and DENV are related
flaviviruses; their homologous proteins and nucleic acids can cause cross-reactions and false-positive results in molecular; antigenic; and serologic diagnostics. We report the
characterization of monoclonal antibody pairs that have been translated into rapid immunochromatography tests to specifically detect the viral nonstructural 1 (NS1) protein antigen and
distinguish the four DENV serotypes (DENV1-4) and ZIKV without cross-reaction. To complement visual test analysis and remove user subjectivity in reading test results; we used image
processing and data analysis for data capture and test result quantification. Using a 30-ml serum sample; the sensitivity and specificity values of the DENV1-4 tests and the pan-DENV test;
which detects all four dengue serotypes; ranged from 0.76 to 1.00. Sensitivity/specificity for the ZIKV rapid test was 0.81/0.86; respectively; using a 150-ml serum input. Serum ZIKV NS1
protein concentrations were about 10-fold lower than corresponding DEN NS1 concentrations in infected patients, moreover; ZIKV NS1 protein was not detected in polymerase chain
reaction- positive patient urine samples. Our rapid immunochromatography approach and reagents have immediate application in differential clinical diagnosis of acute ZIKV and DENV
cases; and the platform can be applied toward developing rapid antigen diagnostics for emerging viruses. © 2017 The Authors; some rights reserved.
Objective: Dengue is a re-emerging public health problem threatening the tropical developing world; mandating rapid diagnosis and supportive management in the absence of licensed
vaccines or anti-dengue therapy. Regions endemic to dengue and related viruses are overwhelmed by the sudden surge of cases during outbreaks. It is difficult to justify confirmatory
diagnosis of every case using The World Health Organization (WHO) criteria or differentiate it from other concurrent viral illnesses. The study evaluated a rapid; sensitive and specific
diagnostic methodology suitable for dengue outbreaks in resource-limited facilities. Methods: There were one hundred dengue patients as per WHO Criteria; as well as 100 healthy controls
from New Delhi; India; were included. Samples collected on the fifth day of onset of fever were tested by lateral flow immunochromatography (LF-ICT); IgM ELISA and reverse transcriptase
polymerase chain reaction (RT-PCR) and results were compared. Diagnostic accuracy indices and Kappa analysis were calculated. Results: The sensitivity; specificity; positive and negative
predictive values (PPV and NPV) of non-structural protein 1 (NS1) against RT-PCR was 98.31%; 100%; 100%; 99.3% and strength of agreement was perfect. Conclusion: Antigen-based and
molecular tests are a better tool for early diagnosis of dengue. The combined LF-ICT kits are highly sensitive; specific; user-friendly; compact; frugal and thus recommended for use in
dengue outbreaks; field conditions and as bed-side diagnostic tests; for confirmatory dengue diagnosis. Further studies are required to assess their utility in prognosis; surveillance and
establishment of guidelines for dengue outbreaks.
OBJECTIVE: To determine the performance of rapid diagnostic tests for dengue and leptospirosis that rely on detecting antibodies that may not be produced when patients present for
medical treatment.; METHODS: We prospectively enrolled 723 patients with undifferentiated febrile illness presenting to rural hospitals in northern and northeastern Thailand over a 1-year
period. We evaluated rapid antibody detection diagnostic tests for dengue and leptospirosis on these patients.; RESULTS: Sensitivity of the tests was low at the acute visit (7.6-21.5%).
Sensitivity at the convalescent visit ranged from 25.8% to 81.5% and was significantly higher than at the acute visit for all tests ([chi]2; P < 0.001).; CONCLUSIONS: Low sensitivity of the
rapid tests at presentation suggests that their utility in the acute phase of dengue and leptospirosis is limited.; Copyright (C) 2007 Blackwell Publishing Ltd.
Background Dengue virus (DENV) infection is prevalent across tropical regions and may cause severe disease. Early diagnosis may improve supportive care. We prospectively assessed the
Standard Diagnostics (Korea) BIOLINE Dengue Duo DENV rapid diagnostic test (RDT) to NS1 antigen and anti-DENV IgM (NS1 and IgM) in children in Cambodia; with the aim of improving the
diagnosis of DENV infection. Methodology and principal findings We enrolled children admitted to hospital with non-localised febrile illnesses during the 5-month DENV transmission
season. Clinical and laboratory variables; and DENV RDT results were recorded at admission. Children had blood culture and serological and molecular tests for common local pathogens;
including reference laboratory DENV NS1 antigen and IgM assays. 337 children were admitted with non-localised febrile illness over 5 months. 71 (21%) had DENV infection (reference assay
positive). Sensitivity was 58%; and specificity 85% for RDT NS1 and IgM combined. Conditional inference framework analysis showed the additional value of platelet and white cell counts
for diagnosis of DENV infection. Variables associated with diagnosis of DENV infection were not associated with critical care admission (70 children; 21%) or mortality (19 children; 6%).
Known causes of mortality were melioidosis (4); other sepsis (5); and malignancy (1). 22 (27%) children with a positive DENV RDT had a treatable other infection. Conclusions The DENV RDT
had low sensitivity for the diagnosis of DENV infection. The high co-prevalence of infections in our cohort indicates the need for a broad microbiological assessment of non-localised febrile
illness in these children. © 2015 Carter et al.
The recent roll-out of rapid diagnostic tests (RDTs) for malaria has highlighted the decreasing proportion of malaria-attributable illness in endemic areas. Unfortunately; once malaria is
excluded; there are few accessible diagnostic tools to guide the management of severe febrile illnesses in low resource settings. This review summarizes the current state of RDT
development for several key infections; including dengue fever; enteric fever; leptospirosis; brucellosis; visceral leishmaniasis and human African trypanosomiasis; and highlights many
remaining gaps. Most RDTs for non-malarial tropical infections currently rely on the detection of host antibodies against a single infectious agent. The sensitivity and specificity of host-

Rapid immunoglobulin M-
based dengue diagnostic test
using surface plasmon
resonance biosensor
Rapid serologic diagnosis of
dengue virus infection using a
commercial capture ELISA that
distinguishes primary and
secondary infections
Rapid tests in the diagnosis of
arboviral diseases
Recombinant envelope
protein-based enzyme
immunoassay for IgG
antibodies is comparable to
neutralization tests for
epidemiological studies of
dengue infection
Recombinant multiepitope
protein for early detection of
dengue infections
Seroprevalence and evaluation
of rapid
immunochromatographic test
and ELISA for the detection of
NS1 antigen; IgM and IgG
antibodies for early diagnosis
of dengue infection in a
Tertiary care hospital; south
India
Seroprevalence of dengue in
Trinidad using rapid test kits: A
cord blood survey
antibody detection tests are both inherently limited. Moreover; prolonged antibody responses to many infections preclude the use of most serological RDTs for monitoring response to
treatment and/or for diagnosing relapse. Considering these limitations; there is a pressing need for sensitive pathogen-detection-based RDTs; as have been successfully developed for
malaria and dengue. Ultimately; integration of RDTs into a validated syndromic approach to tropical fevers is urgently needed. Related research priorities are to define the evolving
epidemiology of fever in the tropics; and to determine how combinations of RDTs could be best used to improve the management of severe and treatable infections requiring specific
therapy. © 2013 The Authors. Clinical Microbiology and Infection © 2013 European Society of Clinical Microbiology and Infectious Diseases.
Surface plasmon resonance (SPR) is a medical diagnosis technique with high sensitivity and specificity. In this research; a new method based on SPR is proposed for rapid; 10-minute
detection of the anti-dengue virus in human serum samples. This novel technique; known as rapid immunoglobulin M (IgM)-based dengue diagnostic test; can be utilized quickly and easily
at the point of care. Four dengue virus serotypes were used as ligands on a biochip. According to the results; a serum volume of only 1â€...μl from a dengue patient (as a minimized volume)
is required to indicate SPR angle variation to determine the ratio of each dengue serotype in samples with 83-93% sensitivity and 100% specificity.
A commercial capture ELISA for specific IgM and IgG antibodies produced during dengue infection (PanBio Dengue Duo) showed excellent sensitivity (99%; n = 78) using sera collected at
hospital discharge compared with established ELISA and hemagglutination inhibition (HAI) assays. Furthermore; the ELISA was able to diagnose 79% of the dengue cases using sera collected
at hospital admission. The ELISA also showed high specificity (92%) in paired sera from patients without flavivirus infection (n = 26); although 45% of the patients with Japanese encephalitis
(n = 20) showed elevation of IgG but not IgM. The IgG capture ELISA showed good correlation with the HAI assay (r = 0.83; P < 0.0001); and IgG levels could be used to distinguish between
primary and secondary infection; with 100% of primary infections and 96% of secondary infections being correctly classified. This ELISA should prove useful in the clinical diagnosis of
dengue infections.
Summary Arboviroses; with in first position dengue fever; threatens millions of people each year; mainly in the intertropical belt. The recent current event was marked by the emergence of
the chikungunya in the New World. Epidemics are sources of imported cases imported in temperate countries. Because of the presence of vector mosquitoes; autochthonous cases of
dengue and chikungunya were described in the South of France. It is the same situation for the West Nile virus infection; which is endemic in temperate regions such as North America.
Rapid diagnosis tests (RDT) are distributed for these three infections. The simple; fast and unitarian format of RDT seems attractive; but numerous studies underline their weakness in terms
of sensitivity. This is added to the classic limit of serologic tests in arboviral diseases; e.g. the delayed appearance of antibodies in the late phase of the acute febrile syndrome. Concerning
dengue; an additional difficulty is the possibility of persistent antibodies from a previous infection. Direct tests of detection of the dengue NS1 antigen under Elisa or RDT format are
available. Their specificity is excellent; but their sensibility is mediocre particularly for RDT. A negative NS1 test does not allow excluding the diagnosis of dengue. A key element of the
interpretation is the delay of sampling with regard to the first day of the clinical signs. Finally; the performances of the arbovirus RDT have to be be analyzed in the light of the
epidemiological context.
Dengue virus (DENV) is the most prevalent arbovirus in the world; found mainly in tropical regions. As clinical manifestations present frequently as nonspecific febrile illness; laboratory
diagnosis is essential to confirm DENV infections and for epidemiological studies. Recombinant envelope (E) antigens of four serotypes of DENV were used to develop an immunoglobulin G
enzyme-linked immunosorbent assay (IgG-ELISA). To evaluate the IgG-ELISA; a panel of serum samples that had been tested previously by a plaque reduction neutralization test (PRNT) was
investigated for the presence of anti-E antibodies against the four DENV serotypes. IgG-ELISA was found to have a sensitivity (91%) and specificity (98%) at a receiver-operating
characteristic (ROC) optimized cutoff and demonstrated high performance as well as good indexes. A concordance of 97% was achieved between both assays; and only 21/704 (3%) samples
were not concordant. The results of the present study demonstrate a moderate correlation between neutralizing antibody titers and IgG-ELISA values. These findings indicate that the
recombinant protein-based IgG-ELISA is a suitable method for routine serodiagnosis; monitoring and seroepidemiological studies of DENV infections. © 2012 Elsevier B.V.
Dengue fever is a mosquito-borne viral disease prevalent mainly in tropical countries. As the clinical manifestations of dengue are not very unique; laboratory diagnosis is crucial in
identifying cases of dengue infection. Detection of dengue infection based on the identification of antidengue antibodies has emerged as a practical and reliable means of diagnosing
dengue fever. We recently developed a customized recombinant dengue multiepitope protein (r-DME-G) that can specifically detect the immunoglobulin G (IgG) class of antidengue
antibodies in patient sera. Using this strategy; we have now created another dengue multiepitope protein; r-DME-;M with specificity for the IgM class of antidengue antibodies. A synthetic
gene encoding the r-DME-M protein was expressed as a maltose-binding protein fusion in Escherichia coli. The recombinant protein was purified in a single affinity chromatographic step to
obtain yields of similar to 15 mg purified protein/liter of culture. The purified protein was used to develop an in-house IgM enzyme-linked immunosorbent assay (ELISA) and tested using a
panel of 172 patient sera characterized using the commercially available Dengue Duo rapid strip test from PanBio; Australia. The IgM ELISA results showed that the r-DME-M protein not
only recognized all IgM(+) samples identified by the PanBio test but also identified samples missed by the latter test. We also successfully adapted the r-DME-M protein to a rapid strip test
format. This approach of creating customized antigens coupled to overexpression in E. coli and simple purification offers a promising alternative option to dengue diagnosis with the
potential to circumvent the drawbacks of the whole virus antigen-based commercial kits.
Dengue is one of the rapidly emerging global threats. In situations of epidemics and routine cases; early diagnosis is the key to successful management of dengue cases. There are many
diagnostic kits available commercially;but their validity is unknown. The standard test is ELISA. In the present study; the test results of rapid immunochromatographic test is compared with
ELISA. Materials & Method: The study was conducted from December 2012 to August 2014 at Shri B M Patil Medical College & Research centre; Bijapur; Karnataka. Probable dengue cases
were diagnosed as per the WHO criteria and rapid immunochromatographic test and ELISA were performed on the serum samples for the detection of NS1 antigen; IgM & IgG antibodies.
Results were analyzed. Results: A total of 90 cases were enrolled. 36 (40%)cases were found to be positive by rapid immunochromatographic test; whereas 39(43%) cases were found to be
positive by ELISA. The sensitivity of rapid test was 92.31% & specificity was 100% along with positive predictive value of 100% and negative predictive value of 94.44% and was compared
with other studies. Conclusion: The study showed that the rapid test can be used as a screening test. Highly suspicious cases should be subjected to investigations with higher sensitivity &
specificity(ELISA); though the results take more time. © 2016; Indian Journal of Public Health Research and Development. All rights reserved.
A cross-sectional sero-epidemiological study was conducted to determine the prevalence of dengue in Trinidad. Two commercial rapid test kits; PanBio Dengue Duo IgM and IgG Rapid Strip
Test and the Bio-Check Plus Dengue G/M Cassette Test (Brittney) were used. The immunosorbent assay (ELISA) (FOCUS Technologies; California) was used as the control. One hundred and
twenty five cord blood samples were collected (46 from Mt. Hope Women's Hospital (MH) and 79 from the San Fernando General Hospital (SF)). All blood samples were tested in
accordance with the two rapid kits and ELISA assay manufacturer's instructions. From 125 cord blood samples; the IgG FOCUS ELISA results showed 93.5 and 95% infections at MH and SF;
respectively. Whereas the Brittney and PanBio kits showed 10.9 and 5.1%; and 26.1 and 50.6% for MH and SF; respectively. Based on the FOCUS ELISA (control) assays; the combined
seroprevalence rate from north and south Trinidad was 94.4%. IgG and IgM sensitivity and specificity levels were higher in the PanBio than Brittney test kits. The high seroprevalence rates
observed in Trinidad are discussed to stimulate more research to explain this phenomenon and to prevent the Southeast Asian scenario from developing in the Americas. (c) 2007 Elsevier
B.V. All rights reserved.

Temperature and the field
stability of a dengue rapid
diagnostic test in the tropics
Temperature of a Dengue
Rapid Diagnostic Test under
Tropical Climatic Conditions: A
Follow Up Study
The clinical; serological and
molecular diagnosis of
emerging dengue infection at
a tertiary care institute in
Southern; India
The diagnostic sensitivity of
Dengue Rapid test assays is
significantly enhanced by
using a combined Antigen and
Antibody testing approach
The performance of the SD
BIOLINE Dengue DUO® rapid
immunochromatographic test
kit for the detection of NS1
antigen; IgM and IgG
antibodies during a dengue
type 1 epidemic in Jamaica
Two cases of false-positive
dengue non-structural protein
1 (NS1) antigen in patients
with hematological
malignancies and a review of
the literature on the use of
NS1 for the detection of
dengue infection
Use of a rapid test for
diagnosis of dengue during
suspected dengue outbreaks
in resource-limited regions
The global incidence of dengue has increased significantly in recent decades; resulting in a large public health burden in tropical and subtropical countries. Dengue rapid diagnostic tests
(RDTs) can provide accurate; rapid accessible diagnosis for patient management and may be easily used by health workers in rural areas. However; in dengue-endemic areas; ambient
temperatures are often higher than manufacturer's recommendation. We therefore evaluated the effect of high temperature over time on the performance of one commonly used dengue
RDT; the Standard Diagnostics Bioline Dengue Duo. RDTs were kept in five different conditions (at 4°C; 35°C; 45°C; 60°C; and at fluctuant ambient temperatures in a free-standing hut) for
between 2 days and 2 years in the Lao People's Democratic Republic (PDR). RDTs were tested with four control sera (negative; dengue nonstructural protein 1 [NS1]; anti-dengue
immunoglobulin [Ig] ;M and antidengue IgG positive). The RDTs had 100% consistency over the 2-year study; despite high temperatures; including in the hut in which temperatures
exceeded the manufacturer's recommendations for 29% of time points. These data suggest that the diagnostic accuracy of the SD Bioline Dengue Duo RDT remains stable even after long-
term storage at high temperatures. Therefore; use at such ambient temperatures in tropical areas should not jeopardize the dengue diagnostic outcome. Copyright © 2015 by The
American Society of Tropical Medicine and Hygiene.
The Dengue Duo Rapid Diagnostic Test (SD Dengue RDT) has good specificity and sensitivity for dengue diagnosis in rural tropical areas. In a previous study; using four control sera; we
demonstrated that that the diagnostic accuracy of these RDTs remains stable after longterm storage at high temperatures. We extended this study by testing sera from 119 febrile patients
collected between July-November 2012 at Salavan Provincial Hospital (southern Laos) with RDTs stored for 6 months at 4°C; 35° and in a hut (miniature traditional house) at Lao ambient
temperatures. The dengue NS1 antigen results from RDTs stored at 35°C and in the hut demonstrated 100% agreement with those stored at 4°C. However; lower positive percent
agreements; with broad 95%CI; were observed for the tests: Ig;M 60% (14.7-94.7) and 40% (5.3-85.3) for RDTs store at 35°C and in the hut; compared to those stored at 4°C; respectively.
This study strenghtens the evidence of the robustness of the NS1 antigen detection RDT for the diagnosis of dengue after storage at tropical temperatures. © 2017 Sengvilaipaseuth et al.
This is an open access article distributed under the terms of the Creative Commons Attribution License; which permits unrestricted use; distribution; and reproduction in any mediu;M
provided the original author and source are credited.
Introduction: Dengue is an acute viral infection which presents as uneventful pyrexia to a fatal complication. This infection is increasingly being recognized as the world's major emerging
tropical disease and an important public health problem. This article highlights the clinical manifestations of Dengue virus infection and the various molecular tests that were used for its
laboratory diagnosis. Methods: Serum samples from 713 suspected cases of Dengue were collected between August and December 2007. The clinical profiles of 123 hospitalized patients
were analyzed. Serology; RT- PCR; virus isolation and sequencing were done. Results: The most common clinical symptoms were fever; thrombocytopenia; rash and elevated liver enzymes.
The demonstration of the Dengue RNA in 5.16% samples; the detection of Dengue specific IgM antibodies in 18% samples and the isolation of the DENV-4 and the DENV-3 viruses from the
clinical samples confirmed this Dengue outbreak. A co-infection with Chikungunya was observed in 2.06% of the cases. The phylogenetic analysis revealed that the Indian Dengue-4 isolates
from this outbreak belonged to the genotype I. This study clearly indicated the sudden dominance of DENV-4 in an Indian Dengue outbreak. Conclusion: The surveillance of the Dengue
viruses needs to be closely monitored for the emergence of newer serotype(s) in hitherto unknown areas.
Background: Serological tests for IgM and IgG are routinely used in clinical laboratories for the rapid diagnosis of dengue and can differentiate between primary and secondary infections.
Dengue virus non-structural protein 1 (NS1) has been identified as an early marker for acute dengue; and is typically present between days 1-9 post-onset of illness but following
seroconversion it can be difficult to detect in serum. Aims: To evaluate the performance of a newly developed Panbio® Dengue Early Rapid test for NS1 and determine if it can improve
diagnostic sensitivity when used in combination with a commercial IgM/IgG rapid test. Methodology: The clinical performance of the Dengue Early Rapid was evaluated in a retrospective
study in Vietnam with 198 acute laboratory-confirmed positive and 100 negative samples. The performance of the Dengue Early Rapid in combination with the IgM/IgG Rapid test was also
evaluated in Malaysia with 263 laboratory-confirmed positive and 30 negative samples. Key Results: In Vietnam the sensitivity and specificity of the test was 69.2% (95% CI: 62.8% to 75.6%)
and 96% (95% CI: 92.2% to 99.8) respectively. In Malaysia the performance was similar with 68.9% sensitivity (95% CI: 61.8% to 76.1%) and 96.7% specificity (95% CI: 82.8% to 99.9%)
compared to RT-PCR. Importantly; when the Dengue Early Rapid test was used in combination with the IgM/IgG test the sensitivity increased to 93.0%. When the two tests were compared
at each day post-onset of illness there was clear differentiation between the antigen and antibody markers. Conclusions: This study highlights that using dengue NS1 antigen detection in
combination with anti-glycoprotein E IgM and IgG serology can significantly increase the sensitivity of acute dengue diagnosis and extends the possible window of detection to include very
early acute samples and enhances the clinical utility of rapid immunochromatographic testing for dengue. © 2011 Fry et al.
Background: Dengue is an important mosquito-borne viral infection that affects millions of persons worldwide. Early diagnosis is necessary to effect appropriate management and decrease
mortality. Immunochromatographic tests are advantageous in producing dengue test results within 30 min but these results should be sensitive and specific. In this study we evaluated the
diagnostic performance of the SD BIOLINE Dengue DUO® rapid immunochromatographic test kit. A panel of 309 dengue and 30 non-dengue single serum samples characterized by using
reference enzyme-linked immunosorbent assays (ELISAs) was used. These samples were received in the virology laboratory for routine testing during a dengue type 1 outbreak between
October to December; 2012. Results: The overall diagnostic sensitivities of the SD BIOLINE Dengue DUO® rapid testfor Ig;M IgG and NSI were 49.3 % (95 % CI: 41.3-57.4); 39.1 % (95 % CI:
33.3-45.2) and 90 % (95 % CI: 82.1-94.7); respectively. The IgM and IgG detection rates were significantly lower than that of the NSI (p < 0.001). However the combination of the IgM
detection with NS1 detection or both NS1 and IgG resulted in a significant (p < 0.001) increase in sensitivity to 97.5 % (95 % CI: 92.9-99.2) and 98.9 % (95 % CI: 96.0-99.7); respectively.
These higher sensitivities were achieved without any decrease in specificities. Conclusions: This study revealed that combining two or more parameters of the SD BIOLINE Dengue DUO®
rapid kit significantly improved the sensitivity of diagnosis of dengue virus infection and supports its usefulness in the Jamaican setting. © 2015 Vickers et al.
Early diagnosis of dengue has been made easier in recent years owing to the advancement in diagnostic technologies. The rapid non-structural protein 1 (NS1) test strip is widely used in
many developed and developing regions at risk of dengue. Despite the relatively high specificity of this test; we recently encountered two cases of false-positive dengue NS1 antigen in
patients with underlying hematological malignancies.We reviewed the literature for causes of falsepositive dengue NS1. Copyright © 2015 by The American Society of Tropical Medicine
and Hygiene.
Dengue is major public health proble;M globally. Timely verification of suspected dengue outbreaks allows for public health response; leading to the initiation of appropriate clinical care.
Because the clinical presentation of dengue is nonspecific; dengue diagnosis would benefit from a sensitive rapid diagnostic test (RDT). We evaluated the diagnostic performance of an RDT
that detects dengue virus (DENV) nonstructural protein 1 (NS1) and anti-DENV IgM during suspected acute febrile illness (AFI) outbreaks in four countries. Real-time reverse transcription-
PCR and anti-DENV IgM enzyme-linked immunosorbent assay were used to verify RDT results. Anti-DENV IgM RDT sensitivity and specificity ranged from 55.3 to 91.7% and 85.3 to 98.5%;
respectively; and NS1 sensitivity and specificity ranged from 49.7 to 92.9% and 22.2 to 89.0%; respectively. Sensitivity varied by timing of specimen collection and DENV serotype. Combined

Use of recombinant envelope
proteins for serological
diagnosis of Dengue virus
infection in an
immunochromatographic
assay.
Validation and reliability of
the rapid diagnostic test ‘SD
bioeasy dengue duo’ for
dengue diagnosis in Brazil: A
phase III study
Virological surveillance of
dengue in Saint Martin and
Saint Barthélemy; French West
Indies; using blood samples on
filter paper
test results moderately improved the sensitivity. The use of RDTs identified dengue as the cause of AFI outbreaks where reference diagnostic testing was limited or unavailable. Copyright ©
2016 David et al.
An immunochromatographic test that incorporates recombinant antigens (Dengue Duo Rapid Strip Test, PanBio; Brisbane; Australia) has recently become commercially available. This
assay is performed in 15 min and detects both immunoglobulin M (IgM) and IgG in a capture format. The four recombinant proteins used represent the N-terminal 80% of the viral
envelope glycoproteins of dengue viruses 1; 2; 3; and 4; respectively. The sensitivity and specificity of the recombinant-antigen-based assay were 90 and 86%; respectively. The similar
diagnostic performance of these antigens to that of enzyme-linked immunosorbent assays using whole dengue virus suggests that they mimic whole dengue viruses in primary structure
and epitope conformation. These results suggest that recombinant proteins can be used in diagnostic assays for dengue to overcome safety issues associated with the use of whole virus.
BACKGROUND The diagnosis of dengue is complex. Until recently; only specialised laboratories were able to confirm dengue infection. However; this has changed with the newly available
immunochromatographic rapid tests. Early diagnosis is of great interest; and point-of-care rapid tests have been increasingly used in Brazil. Most of those tests have not undergone
validation in the Brazilian population. In this context; we decided to evaluate a rapid test introduced in the Federal District (FD). OBJECTIVES To estimate the accuracy and reliability of the
SD Bioeasy Dengue Duo rapid test and its components to detect dengue infections in a consecutive sample of symptomatic residents in the FD; Brazil. METHODS In total; 1353 venous blood
samples were collected between 2013 and 2014. Two hundred and six positive samples (cases) and 246 negative samples (non cases) were required for sensitivity and specificity
estimation; respectively, for agreement evaluation; we used 401 samples. The reference standard used was a composite of MAC-ELISA; virus isolation and real-time polymerase chain
reaction (RT-qPCR). The evaluation was conducted prospectively under field conditions in the public health units of the FD. FINDINGS The results for the overall accuracy of the rapid test
(NS1/IgM combined) showed 76% sensitivity and 98% specificity. The sensitivity for the NS1 component (67%) was better than that for the IgM component (35%). The positive likelihood
ratio was 46; and the negative likelihood ratio was 0.24. The reliability of the test (NS1/IgM combined) demonstrated crude agreement of 98% (Kappa index 0.94). MAIN CONCLUSIONS The
present phase III; large-scale validation study demonstrates that the rapid test SD Bioeasy Dengue Duo has moderate sensitivity (NS1/IgM combined) and high specificity. Therefore; the
test is useful in confirming the diagnosis of dengue; but not enough to rule out the diagnosis. Our results also suggest that Dengue virus (DENV) viral load estimated through the RT-qPCR
and antibody level measured through the MAC-ELISA could have had a direct influence on the accuracy of the rapid test. © 2018; Fundacao Oswaldo Cruz. All rights reserved.
To strengthen active dengue surveillance in Saint Martin and Saint Barthélemy; two French Caribbean islands; we evaluated the epidemiological usefulness of collecting blood samples from
NS1-positive dengue patients on filter paper to identify the dengue serotypes circulating in these regions during a 27-month period. This approach allowed dengue serotypes to be
identified by reverse transcriptase-polymerase chain reaction in 90.1% of the total set of 666 samples analyzed and; in 95.5% of the samples collected during the acute phase of the disease.
This prospective virological surveillance using blood samples absorbed onto filter paper; which were stored at 4°C and shipped at ambient temperature to a specialized laboratory for
analysis; allowed us to avoid the logistic and financial costs associated with shipping frozen venous blood samples. This surveillance system offers a low-cost alternative for reinforcing
dengue prevention in areas where specialized laboratories do not exist; notably by facilitating the early detection of potentially new dengue serotypes. Copyright © 2012 by The American
Society of Tropical Medicine and Hygiene.