African Swine Fever Virus antibody Blocking ELISA (b-ELISA) Kit for pig
- Sep 09, 2026 Anon
African Swine Fever Virus antibody Blocking ELISA (b-ELISA) Kit
Catalog No. LSY-30048
1.Usage
This kit is used to detect ASFV antibodies in serum and plasma of pigs, evaluate the immune status of ASFV vaccines in pig farms, and assist in serological diagnosis of infected pigs.
2.The kit components
1 | ASFV antigen coated microplate | 96TX2 |
2 | ASFV Negative control serum | 2 ml |
3 | ASFV Positive control serum | 2 ml |
4 | Enzyme conjugate concentrate | 0.24 ml |
5 | Enzyme conjugate dilution | 24 ml |
6 | Sample diluent | 12ml |
7 | Substrate | 24 ml |
8 | Stop solution | 15 ml |
9 | 25×concentrated washing buffer | 50 ml |
10 | Cover film | 2 pieces |
11 | Instruction | 1 piece |
3. Test procedure
3.1. Reagent preparation
3.1.1 Washing buffer: Before use, the 25×concentrated washing buffer should return to room temperature (20-25℃) and shaken to dissolve the precipitate (preferably heated in a 37 ℃ constant temperature incubator for 5-10 minutes). Then, it should be diluted 25 times with deionized water (e.g. 30 mL of concentrated washing solution mixed with 720 mL of deionized water) and mixed well. The diluted washing buffer can be stored at 2-8℃ for 7 days.
3.1.2 Preparation of Enzyme Conjugate Working Solution: Before use, the Enzyme conjugate concentrate should return to room temperature (20-25℃), then dilute it with Enzyme conjugate dilution at 1:99, (e.g. 0.01mL of Enzyme conjugate concentrate +0.99mL of Enzyme conjugate dilution) and set aside for immediate use.
Note: The preparation volume should be determined based on the actual quantity of samples.
3.2 Sample Processing
Take animal whole blood, wait for the blood to coagulate, centrifuge at 4000 rpm for 10 minutes, and collect the supernatant. It can also naturally coagulate and precipitate the serum;
Note: Serum should have no hemolysis.
3.3. Procedure
3.3.1 All reagents should return to room temperature (20-25℃) before use.
3.3.2 Take the ASFV antigen coated microplate (which can be disassembled and divided according to the sample size), add 50μL Sample diluent into each well. Set up negative and positive controls each 2 wells, and take 50μL of negative and positive control serum and add them to their wells accordingly. Add 50μL of serum to the rest reaction wells. Gently shake the sample in the well, cover it with a cover film, and react at 37℃ in the dark for 60 minutes.
3.3.3 Discard the solution from the well, add 300μL of diluted washing buffer to each well, let it stand for 30 seconds, and then discard the washing buffer. Wash 4 times and pat dry on absorbent paper. Note: Do not use equipment for centrifugal spin drying.
3.3.4 Add 100μL of enzyme conjugate working solution(currently prepared) to each well, cover with a cover film, and react at 37℃ in the dark for 30 minutes.
3.3.5 Discard the solution from the well and wash 4 times using the same method as in 3.3.3.
3.3.6 Add 100μL of Substrate to each well, cover with a cover film, and react at 25±2℃ in the dark for 10 minutes.
3.3.7 Add 50μL of Stop solution to each well and measure the results within 5 minutes.
3.3.8 Use an enzyme-linked immunosorbent assay (ELISA) reader to read the OD values of each well at 450 nm (630 nm as the reference wavelength).
4. Result judgement
For the assay to be valid, the average OD value of Negative control wells/ the average OD value of Positive control wells≥3. Otherwise the test is invalid, need test again.
Calculate the blocking rate (%) as following:
X%= [(Average OD value of Negative control - OD value of Sample)/ (Average OD value of Negative control- Average OD value of Positive control)]x100%
If X%≥50%: it is ASFV antibody positive;
X%＜40%: it is ASFV antibody negative;
The sample test result is between the two, indicating a suspicious sample
5. Precautions and warnings for users
(1) Before using, each reagent should return to room temperature. Store back at 2-8 ℃ after use.
(2) Different batches of kit components should not be mixed, and cross contamination should be prevented when using different reagents.
(3) Substrate and Stop solution should not be exposed to strong light or in contact with oxidants.
(4) When diluting 25×concentrated washing buffer, if crystals are found, they should be heated to dissolve before use.
(5) When pipetting, it should be done as accurately as possible to prevent the formation of bubbles.
(6) The time and temperature specified in each operating procedure should be strictly followed.
(7) The solid and liquid waste generated after the use of this product should be disposed of in accordance with local regulations to avoid pollution.
Specifications: 96*2 wells.
Expiry date: 12 months.
Storage: Store at 2~8℃, in the dark, no frozen.
Production Date: On outer-packing of the test kit, use the kit during expiry date.
For veterinary diagnostic use only
Shenzhen Lvshiyuan Biotechnology Co., Ltd
D Building, National Biological Industrial Park of Marinelife, No.2 Binhai Road, Dapeng, Shenzhen, 518120 China
Tel. 86-755-28438788/86-18165709090 Fax 86-755-28938800
Email: info@lsybt.com www.lsybt.com
