Development and Optimization of Indirect ELISA Detection Method of Classical Swine Fever Virus Antibody |
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| Authors: | FENG Chun-hua ZHU Yan-ping GUO Dong-guang YUE Feng |
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| Institution: | 1. Xinxiang Animal Epidemic Prevention and Control Center, Xinxiang 453003, China;2. Biotechnology Research Center, Xinxiang University, Xinxiang 453003, China |
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| Abstract: | In this study,through a series of screening and optimization of reactions condition,an indirect ELISA method was developed for detections the antibodies against classical swine fever virus (CSFV) with the purified recombinant CSFV E2 protein.The results showed that the optimal concentration for coating antigen was 1.0 μg/mL and incubated at 37 ℃ for 1 h.The proper serum sample was diluted by 1:200 and the sealing time was 37 ℃ for 1 h.Enzyme labeled second antibody was diluted by 1:10 000 and the reaction time was 45 min incubating at 37 ℃.The D450 nm<0.30 of sample defined as negative of CSFV antibody in the serum sample.The intra-batch and inter-batch variation coefficients were 4.8% and 6.9%,respectively.It indicated that the method in this study had a high stability.Specific test showed that the indirect ELISA had no crossing reactions with the antibodies against PRV,PRRSV,PCV and PPV.80 serum samples were detected in this study,the positive coincidence rate of indirect ELISA was 83.58%,the negative coincidence rate was 76.92%,and the total coincidence rate was 80.25% compared to the results of import blocking ELISA kit.The results suggested that the established indirect ELISA method in this study had an excellent clinical application. |
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| Keywords: | classical swine fever virus (CSFV) E2 protein indirect ELISA antibody detection |
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