&nbsp;Establishment and Optimization of Indirect ELISA for Detection of
 Structural Protein E0 Antibody of Classical Swine Fever Virus

GUO Dongguang, CHEN Mingyan, ZHU Yanping, LI Peng, YUE Feng, CUI Fangwei, LI Wenming, WANG Jinhai, WANG Xuannian

doi:10.15933/j.cnki.1004-3268.2019.11.021

Journal of Henan Agricultural Sciences >

2019 , Vol. 48 >Issue 11: 151 - 156

DOI: https://doi.org/10.15933/j.cnki.1004-3268.2019.11.021

Animal Husbandry and Veterinary Medicine

Establishment and Optimization of Indirect ELISA for Detection of Structural Protein E0 Antibody of Classical Swine Fever Virus

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( 1.School of Life Science,Xinxiang University,Xinxiang 453003,China; 2.School of Life Science,Zhengzhou University,Zhengzhou 450000,China)

Received date: 2019-06-15

Online published: 2019-11-15

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Abstract

In order to establish an economical and reliable method for the detection of serum antibody against classical swine fever virus(CSFV),the recombinant protein E0 expressed in Escherichia coli was used as the coating agent，an indirect ELISA(E0-ELISA) was developed for the detection of serum antibody against CSFV.The results showed that the optimized E0-ELISA best effect was gained when the E0 protein was coated at 37℃ for 1 h and the protein mass concentration at 1.0 μg/mL.The best blocking condition was 5% skimmed milk powder solution at 37℃for 2 h.The best effect was obtained when the serum to be tested was diluted at 1∶ 100 and incubated at 37 ℃ for 60 min,and when the second antibody was diluted at 1∶ 5 000 and incubated at 37℃ for 45 min.When OD450＜0.370,it was negative for CSFV antibody.The variation coefficients of intra-batch and inter-batch repeated tests were 1.99%—7.69% and 2.16%—9.23%,respectively,which showed a good stability for the indirect ELISA.The positive serum of PＲＲSV,PＲV,PCV and PPV tested by E0-ELISA showed the negative results.It was still positive when the positive serum CSFV was diluted to 1∶ 640,showing that E0-ELISA had a good sensitivity.Compared with IDEXX CSFV antibody detection kit，the E0-ELISA coincidence rate,sensitivity and specificity were 92.00%,97.86% and 78.33%,respectively.All the results indicated that an economical,stable,specific and sensitive indirect ELISA for detection of CSFV antibodies was successfully established．

Key words： CSFV; Structural protein E0; Antibody detection; Indirect ELISA

Cite this article

GUO Dongguang, CHEN Mingyan, ZHU Yanping, LI Peng, YUE Feng, CUI Fangwei, LI Wenming, WANG Jinhai, WANG Xuannian . Establishment and Optimization of Indirect ELISA for Detection of Structural Protein E0 Antibody of Classical Swine Fever Virus[J]. Journal of Henan Agricultural Sciences, 2019 , 48(11) : 151 -156 . DOI: 10.15933/j.cnki.1004-3268.2019.11.021

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