摘要
为建立PRRSV抗体的检测方法,通过扩增PRRSV ORF6基因,将其克隆至pET-28a(+)中,IPTG诱导重组质粒转化菌表达,经SDS-PAGE和Western blot鉴定,表明重组质粒能够表达目的蛋白;经亲和层析纯化表达蛋白,以SPA标记胶体金,用纯化M融合蛋白和羊抗猪抗体为检测线和质控线研制胶体金试纸,检测猪血清样品,比较与IDEXX ELISA试剂盒检测结果的符合率。结果表明,表达蛋白具有良好的反应性,制备的胶体金试纸与IDEXX ELISA试剂盒检测符合率为89%,该方法为PRRSV抗体的检测提供了快速简便的方法。
In order to establish a method for detection of PRRSV antibodies, the ORF6 gene was amplified and inserted into pET-28a vector. The recombinant plasmid transformed bacteria was induced by IPTG. Identification by SDS-PAGE and Western blot showed that the target gene was expressed correctly. The expressed product was purified by affinity chromatography and used to develop the method of colloidal gold strip for detecting antibodies against PRRSV. The serum samples were tested by colloidal gold strip and was compared with IDEXX ELISA Kit. The results showed that the expression protein had good reactivity. The coincidence rate of the colloidal gold strip to IDEXX ELISA kit was 89%.The established method was rapid and simple for detecting PRRSV antibodies.
出处
《中国兽药杂志》
2014年第1期25-28,共4页
Chinese Journal of Veterinary Drug
基金
吉林省世行贷款农产品质量安全项目(2011-Y28)
科技部农业科技成果转化资金项目(2012GB2B100106)
关键词
猪繁殖与呼吸综合征病毒
表达
胶体金试纸
porcine reproductive and respiratory syndrome virus
non glycosylated membrane protein M
expression
colloidal gold strip
