山羊痘病毒P32基因的截短表达及间接ELISA方法的建立和应用  

Establishment and application of an indirect ELISA for prokaryotic expression of truncated P32 protein of goatpox virus

作　　者：孟卫芹 王金良[1] 许崇友 陈金龙 沈志强[1] MENG Weiqin;WANG Jinliang;XU Chongyou;CHEN Jinlong;SHEN Zhiqiang(Shandong Binzhou Animal Science&Veterinary Medicine Academy,Binzhou,Shandong 256600,China;Rizhao Donggang Ani mal Disease Control Centre,Rizhao,Shandong 276800,China;College of Animal Science and Technology,Shihexi University,Shihexi,Xinjiang 832003,China)

机构地区：[1]山东省滨州畜牧兽医研究院,山东滨州256600 [2]日照市东港区动物疫病预防控制中心,山东日照276800 [3]石河子大学动物科技学院,新疆石河子832003

出　　处：《中国兽医学报》2022年第9期1775-1779,共5页Chinese Journal of Veterinary Science

基　　金：山东省现代农业产业技术体系羊创新团队岗位专家基金资助项目(SDAIT-10-07)。

摘　　要：为建立一种山羊痘病毒(GTPV)抗体快速检测方法,通过PCR技术扩增了P32蛋白优势抗原区(2-141AA)基因序列,将其连接至原核表达载体pET-28a(+)上,转化至E.coli BL21(DE3)中,经IPTG诱导后获得了重组蛋白,纯化后经Western blot鉴定表明该蛋白具有良好的特异性和抗原反应性。以重组蛋白作为包被抗原,通过优化反应条件建立了快速检测GTPV抗体的间接ELISA方法,该方法具有良好的特异性、敏感性与可重复性,为GTPV的免疫抗体检测及流行病学调查提供了一种血清学诊断方法。in order to establish a rapid method for detecting goatpox virus(GTPV)antibodies,the dominant antigen region(2141AA)of P32 protein gene was amplified by PCR,it was cloned into prokaryotic expression vector pET-28a(+),transformed into E.coli BL21(DE3),the recombinant protein was obtained by IPTG induction.Western blot showed that the protein had good specificity and antigenic reactivity.Using the recom binant protein as coating antigen,an optimized indirect ELISA method was established to detect GTPV antibody.This method had good specificity,sensitivity and repeatability,it provides a new serological diagnosis method for GTPV immune antibody detection and epidemiologic investigation.

关 键 词：山羊痘病毒 P32基因 截短表达 间接ELISA 

分 类 号：S852.65[农业科学—基础兽医学]