The Biokau® hog cholera virus antibody test kit, or called
Classical Swine Fever Virus Antibody ELISA Test Kit (CSFV Ab), is
used for the detection of hog cholera virus antibodies in porcine
serum; assessment of immunity conditions against hog cholera virus
in the pig farms and epidemiology investigation of the hog cholera.
The Biokau® hog cholera virus antibody 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 hog cholera virus in porcine serum. In the test,
the coated antigen combine with CSFV-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
CSFV-IgG, when the value of color is greater than the cut-off
value, the pigs are vaccinated well.
1 CSFV coated microplate 2 pieces (96 wells)
2 CSFV Negative serum(green lid) 1 tube (1 mL)
3 CSFV Positive serum (red lid) 1 tube (1 mL)
4 CSFV Enzyme conjugate (yellow lid) 1 bottle (22 mL)
5 20×Wash Concentrate (white lid) 1 bottle (50 mL)
6 Substrate A (red lid)/B (black lid) 1 bottle each (12 mL/bottle)
7 Stop solution(blue lid) 1 bottle (12 mL)
8 Sample diluent solution(transparent lid) 1 bottle (25 mL)
9 Adhesive Foil 6 pieces
10 Instruction 1 piece
4. Material Required But not Provided
- Microplate Reader(wave length: 450/630 nm).
- Microplate Washer.
- Micropipettes, adjustable.
- Constant temperature box or incubator.
5. Sample requirement
- 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℃.
- Avoid to use the samples with severe hemolysis, precipitate,
contaminated by bacteria or protein suspension.
- The EDTA, heparin sodiun and other anticoagulants will not affect
- Take out the coated plates(Can be detached) and record the sample
position on a worksheet. Set one blank control well, add nothing.
Set 2 wells for negative control serum and 2 wells for positive
control serum, 100 μL/well. Others are wells for samples, add
100μL/well of Sample diluent solution, then add 10 μL serum sample
- Mix gently, incubate at 37℃ for 30 min.
- Remove adhesive foil. Pour the liquid out of the wells, add Washing
solution(dillute the Wash Concentrate at 1:19 with distilled water)
into each well, 300 μL/well, stand for 1 min. Repeat 5 times, at
last time pat to dry on absorbent paper.
- Add 100 μL enzyme comjugate into each well(except the blank well).
- Cover plate with new adhesive foil. Incubate at 37 ℃ for 30 min.
- Repeat step 3(washing).
- Add substrate A one drop (50 μL) and substrate B one drop (50 μL)
into each well, mix properly, incubate for 15 min at 37 ℃ in the
dark with new adhesive foil.
- Add stop solution one drop (50 μL) into each well, mix gently and
determine the result within 5-30 min.
- Measure the optical density(OD) with a photometer at 450
nm(Reference-wavelength: 630 nm). Set zero for the blank well, and
read the OD 450 value of samples on the Microplate reader.
7. ELISA analysis
For the assay to be valid the following specifications must be met.
The CSFV-Positive control mean (PC:OD450) must be greater than
0.70, the CSFV-Negative control mean must be less than 0.05.
Cut-Off Value (C.O.) = 0.2×PC:OD450. The presence or absence of
antibody to CSFV is determined by calculating the sample to C.O.
If the S/C.O. ≥1, the sample is classified as POSITIVE for CSFV
If the S/C.O. ＜1, the sample is classified as NEGATIVE for CSFV
Packing : 96 wells×2.
Expiry date: 12 months.
Storage: Storing at 2~8 ℃ in the dark, not freeze.