猪繁殖与呼吸综合征病毒重组N蛋白间接ELISA检测方法的建立及应用

doi:10.16431/j.cnki.1671-7236.2016.11.014

Abstract

Abstract:

To establish a sensitive,specific and efficient method of antibody detection for porcine reproductive and respiratory syndrome virus（PRRSV),PRRSV N protein was expressed through prokaryotic expression system and purified by affinity chromatography to act as a coating antigen.Then an indirect ELISA detection method for serum PRRSV antibody was finally set up after the optimization of reaction conditions.Besides,the research also involved cross reaction,repeated experiments,and comparison with other ELISA kits.It was determined that the optimum concentration of coating antigen was 2 μg/mL and that the dilution ratio for serum was 1:100,with 30 min of incubation and 10 min of chromogenic reaction.With this method,positive serum samples of five common swine pathogens,including classical swine fever virus（CSFV),porcine circovirus（PCV）and porcine pseudorabies virus（PRV）and so on,were tested,and the results were negative.Both intra-assay and inter-assay coefficients of variation were below 10%,and the comparison with commercial ELISA kits indicated that its accuracy was 94.7%.So this indirect ELISA,which had been established in this research,could provide a rapid diagnosis for swine infected by wild PRRSV and applied in epidemiological investigation,as a convenient and efficient serological antibody detection method.

Key words: porcine reproductive and respiratory syndrome virus（PRRSV); N protein; ELISA

CLC Number:

S858.28

CHENG Fu-liang, LI Fu-hui, LIU Yang-qing, NIE Zhao-jing, CHEN Tian-tian, FANG Dong, FAN Mei-na, GU Wei, WANG Chun-feng. Establishment and Application of an Indirect ELISA with Recombinant N Protein of Porcine Reproductive and Respiratory Syndrome Virus[J]. , 2016, 43(11): 2900-2906.

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Establishment and Application of an Indirect ELISA with Recombinant N Protein of Porcine Reproductive and Respiratory Syndrome Virus

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