Porcine Reproductive and Respiratory Syndrome Virus (PRRSV) Antibody ELISA kit
Catalog No. LSY-30010
The GreenSpring® Porcine reprodutive and respiratory syndrome virus
antibody test kit (PRRSV Ab) is used for the
detection of PRRSV antibodies in swine serum; assessment of
immunity conditions against porcine reprodutive and respiratory
syndrome virus in the pig farms; and investigation epidemiology of
The GreenSpring® PRRSV Ab ELISA test kit is made from the antigen coated
microtiter plate, goat-anti-pig IgG-HRP and other reagents. It
applies the indirect ELISA principle to test the antibodies against
PRRSV in porcine serum. In the test, the coated antigen
combine with PRRSV-IgG in serum, then add IgG-HRP to specifically
bind with complex of antibody-antigens on the microplate. With
the TMB substrate, it will generate an amount of color. The depth
of color is relative with the content of the PRRSV-IgG.
PRRSV antigen coated microplate
96T X 2
Sample diluent solution
PRRSV-IgG Negative control serum
PRRSV-IgG Positive control serum
20×concentrated washing buffer
4. Material Required not Provided
1 Microplate Reader (Dual-wave length: 450/630 nm).
2 Microplate Washer.
3 Micropipettes, adjustable (Single-wave length
1-100ul,0.5-10ul,multi-wave length 30-300ul).
4 Constant temperature box or water bath box.
6 Disposable tips (10ul, 200ul)
7 Deionized water
5. Sample requirement
1 The samples are porcine serum, which should be collected
with no bacteria. The storage time should be less than 1 week at
2-8 ℃, if for long term, it should be kept at -20℃.
2 Avoid to use the samples with severe hemolysis, precipitate,
contaminated by bacteria or protein suspension.
3 The EDTA, heparin sodiun and other anticoagulants will not
affect the results.
1) Bring ELISA reagents to the room temperature (20-25 ℃)
for 30 min to get best results.
2) Sample dilute: Dilute sample with the sample
diluent solution at 40 times.(There will be color change
after adding solution), the diluted sample need to mix evenly to
get better results.
3) Washing solution preparation: Dilute the 20×concentrated washing
buffer with deionized water at 20 times.
1 Adding sample: Take out the required coated plates according
to sample quantity (Can be detached) and record the sample
position on a worksheet. Set 2 wells for negative control serum and
2 wells for positive control serum, add undiluted negative and
positive control serum to its well accordingly, 100 μL/well.
Others are wells for samples, add 100μL/well of the diluted
Sample. (Can do for both single-well and double-well test).
2 Mix gently for 10s, incubate at 37℃ for 30 min.
3 Remove adhesive foil. Pour the liquid out of the wells, add
Washing solution into each well fully, be static for
about 10s, pour out directly. Repeat 3 times, at last time pat
to dry on absorbent paper.
4 Add 100 μL enzyme comjugate into each well.
5 Cover plate with new adhesive foil. Incubate at 37 ℃ for 30 min.
6 Repeat step 3(washing).
7 Add 100ul substrate into each well, mix properly, incubate for 10 min at 37 ℃ in the dark with new adhesive foil.
8 Add 50 μL stop solution into each well, mix gently
for 10s and determine the result.
9 Measure the OD value with a photometer at 450 nm/630nm.
8. ELISA analysis
Measure the OD Value of each well, for the assay to be valid the
following specifications must be met. The
PRRS-Positive control mean must be equal to or greater than
0.60, the PRRS-Negative control mean must be less than 0.15.
The result is judged by S/P value,
S/P=(Sample OD450/630- NCx)/( PCx- NCx), NCx means Negative control’s average OD450/630 value, PCx means Positive control’s average OD450/630 value
If S/P≥0.2, it is positive; less than 0.2, it is negative.
9. Interpretation of the result
1. Severe hemolysis, fiber protein in the serum separation is not
sufficient, containing erythrocytes, a precipitate, a sample
with bacteria may lead to false positive.
2. Negative results may occur on individual pigs
after vaccines due to individual differences or immune
3. Positive results for serological diagnosis and epidemiological
investigation of swine to be combined with other methods and
10. Limit of test method
PRRSV-IgG can be used as the evaluation of PRRSV vaccine
effect and PRRSV virus infection serological diagnostic
indicators, but can not distinguish the two, if want to distinguish
antibody caused by the vaccine or wild virus infection, it shout be
combined with clinical data, immunization programs etc.
11. Product performance
1. Specificity: use this kit to detect reference serum, the
compliance rate reach 100%.
2. Sensitivity: can reach max 1:5120.
3. Precision: CV(%)no bigger than 8%.
4. Stability: Store at 2℃～8℃ for 12 months or store at 37℃ for
3 days, the result can reach the above 3 standards.
12. Precautions and warnings for users
1. This test kit is suitable for in vitro diagnostics.
2. Do not use reagents expired, do not mix reagents from different
3. Read the Manual carefully before use.
4. Experiment rubbish should be dealt with high pressure steam
sterilization at 121 ℃ for 30 minutes, or treated with
5.0g/L sodium hypochlorite disinfectant for 30 minutes, then
5. MicroWell plate removed from the refrigerated environment should
be balanced moisture to dry at room temperature, then can be
opened. Put back unused MicroWell plate into dry foil bag and
sealed at 4 ℃. Unused liquid reagent should cover caps,
store at 2-8 ℃ in dark with other group components.
6. If the 20×concentrated washing buffer appears crystal, it
is normal, put at 37℃ until been dissolved.
7. Should use Micropipettor to add sample and reagents, and
often proof its accuracy.
8. When adding washing buffer, should be full but no overflow,
avoid appearing free enzyme at mouth of well or cross pollution
9. Stop solution is corrosive, use large amount of water to wash
immediately when touch the skin or clothes.
Packing: 96 wells×2.
Expiry date: 12 months.
Storage: Storing at 2~8 ℃in the dark, not freeze.