犬瘟热病毒重组核蛋白间接ELISA方法的建立  

Establishment of an indirect ELISA with a recombinant nucleocapsid protein as antigen for detection of antibodies against canine distemper virus

作　　者：王琛[1] 袁宝[1] 任文陟[1] 张嘉保[1] 

机构地区：[1]吉林大学农学部实验动物中心

出　　处：《畜牧与兽医》2008年第5期37-40,共4页Animal Husbandry & Veterinary Medicine

基　　金：吉林省科技计划发展项目(20071138)

摘　　要：将已构建成功的重组质粒pGEX-4T-1-N转化大肠杆菌BL21株,在最佳诱导条件下获得犬瘟热病毒(CDV)重组N蛋白。将表达产物纯化后进行SDS-PAGE和W estern-b lot分析,与CDV标准阳性血清呈阳性反应。本研究初步建立了以纯化的N蛋白为包被抗原的间接ELISA检测方法,经初步试验证实,该方法敏感、特异。试验结果表明,大肠杆菌中表达的CDV N蛋白在免疫原性上与天然核衣壳蛋白具有较高相似性,可作为诊断用抗原。The recombinant plasmid pGEX-4T-1-N was transformed into E. coli BL21 and the recombinant nucleocapsid protein of canine distemper virus （CDV） was obtained under the optimized induction condition. The analysis of SDS-PAGE indicated a protein band with the molecular weight of 55ku. Western-blot analysis showed that the recombinant protein had a positive reaction with standard positive blood serum against CDV. The indirect ELISA method was developed using the purified fusion protein of nucleocapsid as antigen. The preliminary clinical application showed that it is a sensitive and specific method in detecting CDV antibodies, and the coincidence rate between the recombinant protein ELISA and IDEXX CDV kit was up to 96.25%.

关 键 词：犬瘟热病毒 核衣壳蛋白基因 原核表达 间接ELISA 

分 类 号：S858.292[农业科学—临床兽医学]