中国兽医科学2018，Vol.48Issue(9)：1086-1092,7.DOI:10.16656/j.issn.1673-4696.2018.0128
猪繁殖与呼吸综合征病毒GP5蛋白间接ELISA抗体检测方法的建立与应用
Establishment and application of an indirect ELISA for the detection of antibodies against GP5 protein of porcine reproductive and respiratory syndrome virus
摘要
Abstract
Porcine reproductive and respiratory syndrome virus(PRRSV)is an important pathogen affecting the world pig industry. The GP5 protein is an important immunoprotective protein encoded by the virus. At present,there are a lot methods developed for detection of anti-PRRSV antibodies,but no commercially ELISA kit is available for the detection of GP5 protein antibodies specially. This study was based on the GP5 gene characteristics of a highly pathogenic PRRSV BB0907 strain,deleting the transmembrane region of the GP5 gene,and the remaining gene fragments were optimized for E. coli biophagy. A prokaryotic expression vector was constructed for the highly expressing recombinant protein of GP5,and the modified GP5 recombinant protein was expressed and purified. And an indirect ELISA method was established by optimizing the reaction conditions. The optimum antigen concentration was 1. 0 μg/mL,the optimal dilution of serum was 1 : 100. It had no cross reaction with the antibodies against porcine circovirus type 2,classical swine fever virus,pseudorabies virus,foot-and-mouth disease virus,encephalomyocarditis virus and parssuis haemophilus, respectively. Compared with the commercial PRRSV ELISA kit,it had the relative sensitivity of 90. 69％ and specificity of 92. 86％. The coincidence rate of the two methods was 91. 11％. The positive rate of PRRSV antibodies in 360 clinical pig serum was 74. 44％. In conclusion, the developed indirect ELISA for the detection antibodies against GP5 protein of porcine reproductive and respiratory syndrome virus in this study has an important application prospect.关键词
猪繁殖与呼吸综合征病毒/GP5蛋白/间接ELISAKey words
porcine reproductive and respiratory syndrome virus/GP5 protein/indirect ELISA分类
农业科技引用本文复制引用
侯贺飞,刘雪威,白娟,宋中宝,姜平..猪繁殖与呼吸综合征病毒GP5蛋白间接ELISA抗体检测方法的建立与应用[J].中国兽医科学,2018,48(9):1086-1092,7.基金项目
国家自然科学基金项目(31672565) （31672565）
国家生猪产业技术体系专项(CARS-36) （CARS-36）
