INTENDED USE

Hardy Diagnostics Saline products are recommended for procedures that require the use of an
isotonic diluent. It is also recommended for use in preparing test suspensions of organisms.

SUMMARY
Saline is useful as a diluent to maintain cell integrity and viability because it lacks properties that
may interfere with biochemical reactions and/or antibiotic susceptibility tests. The concentration
of sodium chloride in 0.85% (normal) Saline provides osmotic protection for microbial cells.
Normal saline is used for preparing microbial suspensions when it is necessary to deliver a set
number of microbes to an identification test battery, to antimicrobial agents, or to growth media
used for disk susceptibility testing. It is also used in the preparing of stock solutions and serial
dilutions of antimicrobial agents. The Clinical and Laboratory Standards Institute (CLSI -
formerly NCCLS) recommends the use of 0.85% Saline to adjust the turbidity of bacterial
suspensions to help maintain cell integrity and viability.(1,2) Saline, 0.45% is the isotonic diluent
recommended for use with the Vitek System.

FORMULA
Ingredients per liter of deionized water:*
Saline, 0.45%:
Sodium Chloride 4.5gm
Saline, 0.85%:
Sodium Chloride 8.5gm
Final pH of both solutions is 6.5 +/- 1.0 at 25C.**
** Final pH of K59 is 5.5-7.0 at 25C which is compatible with the API and Micro ID enteric
bacterial
identification strips.
** Final pH of D12 is 5.5-7.2 at 25C and is consistent with Vitek/bioMeriuex specifications
for this product.
** Final pH of D185 is 5.5-7.2 at 25C.
* Adjusted and/or supplemented as required to meet performance criteria.
STORAGE AND SHELF LIFE
Storage: Upon receipt store at 2-30C away from direct light. Saline should not be used if there
are any signs of deterioration (evaporation, discoloration), contamination, or if the expiration
date has passed. Protect from excessive heat.
The expiration dating on the product label applies to the product in its intact packaging when
stored as directed.

Refer to the document "Storage" on the Hardy Diagnostics Technical Document website for
more information.
Distribution Centers:
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The Hardy Diagnostics manufacturing facility and quality
management system is certified to ISO 13485.
Copyright 1996 - 2016 by Hardy Diagnostics. All rights reserved.
HDQA 2207F Rev. 012816hh
Although it can help diagnose some illnesses, an abnormal ESR does not prove that one has a
certain condition. Other confirmatory tests are always needed. An increased ESR rate may be due
to anaemia, cancers such as lymphoma or multiple myeloma, kidney disease, pregnancy and
thyroid disease. An autoimmune disorder is a condition that occurs when the immune system
mistakenly attacks and destroys healthy body tissue. ESR is often higher than normal in people
with an autoimmune disorder. Common autoimmune disorders include lupus erythematosus and
rheumatoid arthritis in adults or children. Very high ESR levels occur with less common
autoimmune disorders including allergic vasculitis, giant cell arthritis, hyperfibrinogenemia, and
macroglobulinemia - primary necrotizing vasculitis and polymyalgia rheumatica. An increased
ESR rate may be due to some infections including systemic infection, bone infections, infection
of the heart or heart valves, rheumatic fever, severe skin infections such as erysipelas and
tuberculosis. Lower-than-normal levels occur with congestive heart failure, hypofibrinogenemia,
low plasma protein (due to liver or kidney disease), polycythemia and sickle cell anaemia.
Why only sodium citrate is being used as an anticoagulant normally in clinical laboratory for
performing ESR? In order to confirm this, ESR was performed using yet another anticoagulant
EDTA. ESR and C-reactive protein (CRP) are bothmarkers of inflammation5,6. Generally, ESR
does not change as rapidly as does CRP, either at the start of inflammation or as it disappears.
CRP is not affected by as many other factors as is ESR, making it a better marker of
inflammation. CRP is an acute phase protein produced by the liver during an inflammatory
reaction. Since CRP levels in the blood rise more quickly after the inflammatory or infective
process begins, ESR is often replaced with C-reactive protein measurement. There are specific
drawbacks, however, for example, both tests for ESR and CRP were found to be independently
associated with a diagnosis of acute maxillary sinusitis7 so in some cases the combination of the
two measurements may improve diagnostic sensitivity and specificity.
Materials and Methods
This experiment was performed using disposable Westergren tubes. The blood of
OPD of Yenepoya University hospital patients were collected into two sets of non-vaccum
blood collection vials containing sodium citrate
and EDTA respectively. The sodium citrate
International Journal of Pharmacy and Biological Sciences (eISSN: 2230-7605)
Manjula Shantaram * et al Int J Pharm Bio Sci
www.ijpbs.com
Erythrocyte Sedimentation Rate (ESR)
solution, How to perform the test
Principle of ESR
ESR test is based on the principle of sedimentation.
When you left the blood in anticoagulant, undisturbed RBCs will settles down in the
container or Wintrobe tube marked 0 to 100 mm.
Factors influencing ESR
1. Plasma protein
. In normal blood RBCs suspended in the plasma from few aggregates of RBCs.
So rate of sedimentation is slow.
. If there is rouleaux formation that will give false value. So acute phase protein affects
the ESR.
2. Number of RBCs
ESR depends upon the number of RBCs like in anemia decreased number of
RBCs settles down rapidly.
In case of polycythemia RBCs settles slowly and ESR will be low.
3. Shape of RBCs
Changes in the shape affect the ESR like in Sickle cell anemia, ESR may low or zero.
4. Size of the RBCs
Macrocytes cells settles more rapidly than the microcytes because of their large size.

Technical factors affecting ESR, giving false increased value:

1. False increased ESR see in :
2. Test when read after one hours.
3. When there is improper dilution with the anticoagulant.
4. When the tube is not vertical and is tilted to one side.
5. If there is vibration of the tube during test.
6. If blood is not mixed properly or there are microclots.
7. Test done at room temperature >25 C.
Technical factors affecting ESR and giving false lower value:
1. When test is read before one hour.
2. Improper mixing of the blood.
3. Improper dilution of the blood.
4. If blood temperature is low.
5. If there are air bubbles in the tube.
6. If room temperature is <20 C. Labpedia.net
Professor Dr. Riaz Ahmad Bhutta
Dr. Naheed Afroz Syed,
Dr. Asad Ahmad, M.D.