畜牧兽医学报2012，Vol.43Issue(12)：1925-1930,6.
基于狂犬病病毒重组蛋白的ELISA检测方法比较研究
Comparative Study on the Indirect ELISA Methods Based on the Different Recombinant Proteins of Rabies Virus
摘要
Abstract
This study was designed to compare the indirect ELISA methods for detection of dog antibodies against rabies virus based on the recombinant matrix protein (M) and phosphoprotein (P). The M gene of rabies virus LEP-Flury strain was amplified by PCR and cloned into prokary-otic expression vector pGEX-6P-l. The resultant constructed pGEX-RV-M plasmid was transformed into BL21 (DE3) and protein expression was analyzed by SDS-PAGE and Western blot. The results of SDS-PAGE showed that the M protein was efficiently expressed, which were mainly soluble, and purified with the affinity chromatography. The results of Western blot indicated that the recombinant protein M showed good immunogenicity. The indirect ELISA methodwas established with the purified recombinant protein M, and a total of 95 serum samples were detected by the method and the indirect ELISA method based on the recombinant protein P respectively. The results showed that compared with the commercially available ELISA kit coating RV as antigen, the indirect ELISA method based on recombinant protein P had a higher coincidence rate, and it can replace the ELISA method based on RV.关键词
狂犬病病毒/基质蛋白M/原核表达/间接ELISA/磷蛋白PKey words
rabies virus/ matrix protein/ prokaryotic expression/indirect ELISA/ phosphoprotein (P)分类
农业科技引用本文复制引用
程朝飞,宫苗苗,田康乐,王勇,赵占中,史利军,李刚..基于狂犬病病毒重组蛋白的ELISA检测方法比较研究[J].畜牧兽医学报,2012,43(12):1925-1930,6.基金项目
国家高技术研究发展计划(863计划)(2008AA10Z411) （863计划）
北京市科委项目(Z07010501780701) （Z07010501780701）
国家公益行业项目(20083014 （）
200903037-2) （）
