原核表达山羊布氏杆菌M5-90VirB12蛋白及其在检测中的初步应用

Prokaryotic expression of Brucellar melitensis VirB12 protein and primary application in serological detection

为建立山羊布氏杆菌(Brucellar melitensis)血清学检测方法,本实验克隆了B.melitensis M5-90疫苗株的virB12基因,并表达了相应蛋白。经SDS-PAGE和western blot鉴定,重组VirB12蛋白分质量约为25ku,具有较好的抗原性。以纯化的VirB12重组蛋白为抗原建立间接ELISA方法,并检测90份B.melitensis临床血清样品,检测结果与试剂盒及虎红平板凝集试验的检测结果符合率均为91.7%,结果表明,VirB12重组蛋白作为包被抗原可用于B.melitensis感染的血清检测,为进一步建立B.melitensis M5-90virB12缺失标记疫苗的鉴别检测提供方法。

To develop a serological detection method of Brucellar melitensis, we cloned the virB12 gene of B. melitensis and expressed in E. coli. The expressed recombinant VirB12 was 25 ku and reacted specifically with B. melitensis positive serum analyzed by SDS-PAGE and western blot. An indirect ELISA was set up using purified VirB12 as coating antigen. A total of 90 clinic B. melitensis sera were examined and the coincidence was 91.7%, comparing with commercial ELISA Kit and the rose bengal plate test. The VirB12 recombinant protein could be applied for the detection of B. melitensis infection.