应用VirB12蛋白建立检测牛布鲁菌血清抗体的方法  

Detecting sera antibody of bovine brucella by an ELISA method using VirB12 protein

作　　者：王冲[1] 孙勇[1] 戚帅[1] 宋砚泽[1] 田武林[1] 崔敏[1] 薛雨佳[1] 张西臣[1] 闫广谋[1] 

机构地区：[1]吉林大学动物医学学院,吉林长春130062

出　　处：《中国兽医学报》2013年第3期400-403,共4页Chinese Journal of Veterinary Science

基　　金：国家自然科学基金委员会青年科学基金资助项目(31101840);吉林省科技厅科技引导计划青年科研基金资助项目(201101076)

摘　　要：从牛布鲁菌基因组中克隆出VirB12基因,以pET-28α(+)为模板构建VirB12原核表达载体pET-V12,pET-V12在E.coli BL21(DE3)中表达重组VirB12蛋白。经Western-blotting分析,表明重组的VirB12蛋白具有免疫原性。以纯化的VirB12蛋白为抗原,建立了间接ELISA检测牛布鲁菌抗体的方法。经对300份牛血清样品间接ELISA与虎红平板试验(RBPT)检测结果的比较,该方法的敏感性为89%,特异性为98.3%,准确度为92.7%。Cloning VirB12 gene of bovine brucella, constructing VirBl 2 prokaryotic expression vec- tor using pET-28a（＋） as the template,and expressing VirB12 protein in E. coli BL21 （DE3）. The expressed product was analyzed by Western-blotting. The result shows that this protein has immunogenicity to bovine brucella. The purified VirB12 protein was coated as the antigen to con- struct a method of indirect ELISA for detecting bovine brucellosis from sera. Three hundred ser- um samples of bovine were detected by the indirect ELISA method. Its sensitivity was 89 %, speci- ficity was 98.3% and accuracy was 92.7% compared with the rose bengal plate test.

关 键 词：牛布鲁菌 VirB12蛋白 原核表达 间接ELISA 

分 类 号：S852.614[农业科学—基础兽医学]