小反刍兽疫病毒N蛋白原核表达与间接ELISA检测方法的建立  

Prokaryotic Expression of N Protein of Peste des Petits Ruminants Virus and Development of Indirect ELISA

作　　者：孙雨[1] 赵柏林[1] 王晓英[1] 董浩[1] 翟新验[1] 曲萍[1] 胡冬梅[1] 杨天意[1] 石慧[1] 宋晓晖[1] 

机构地区：[1]中国动物疫病预防控制中心,北京102600

出　　处：《动物医学进展》2017年第1期6-10,共5页Progress In Veterinary Medicine

基　　金：国家重点计划研发项目(2016YFD0500901)

摘　　要：小反刍兽疫病毒(PPRV)N蛋白是病毒粒子的主要结构蛋白,也是诊断小反刍兽疫的主要靶蛋白。为获得高效表达的可溶性N蛋白,并建立基于N蛋白的小反刍兽疫的间接ELISA检测方法,通过优化密码子、优化表达条件等条件探索,在大肠埃希菌表达系统中获得了高效表达的可溶性N蛋白,进一步基于N蛋白建立了间接ELISA检测方法,该方法对其他相关的羊类病原无交叉反应,其组内与组间变异系数均低于9%,具有良好的重复性。对480份临床血清样品进行检测,同时用法国IDVET竞争ELISA试剂盒进行比较,符合率达到98.33%。本研究为进一步开发成熟的小反刍兽疫抗体检测试剂盒奠定了基础。Peste des petits ruminants virus （PPRV） N protein is the major structural protein of virus particles,and it is the main target protein in the diagnosis of PPRV.To establish the expression and purification methods for efficient soluble PPRV N protein and indirect ELISA method for diagnosis of PPRV based on N,this study got a high expression of soluble N protein in Escherichia coli expression system through optimization of codon and expression conditions,and established an indirect ELISA detection method based on N protein. The results showed that the assay was specific and repeatable. Furthermore, a total of 480 clinical serum samples were detected by this assay and the agreement was 98.33% with commercial ELISA Kit （IDVET）.This study laid the foundation for the further development of kit for detection of PPRV antibody.

关 键 词：小反刍兽疫 N活性蛋白 可溶性表达与纯化 间接酶联免疫吸附试验 

分 类 号：S852.659.5[农业科学—基础兽医学]