猪圆环病毒2型ORF2基因原核表达蛋白间接ELISA检测方法的建立及应用

Establishment of an indirect ELISA with PCV2 recombinant capsid protein expressed in E.coli and initial application

对含有已构建的猪圆环病毒2型(PCV2)ORF2基因原核表达质粒pET-ORF2s的BL21菌进行诱导表达,纯化重组蛋白包涵体,Western-blot检测纯化蛋白,将其作为抗原包被酶标板,优化间接ELISA试验条件,建立可检测PCV2血清抗体的间接ELISA方法。用该方法对吉林省多个种猪场收集的179份血清样品进行检测,总阳性率为42.5%。由此说明,本试验建立的PCV2血清抗体间接ELISA方法,具有较高的敏感性和特异性,适用于大规模猪场血清学检测。

The BL21 E.coli with recombinant plasmid pET-ORF2s of porcine circovirus type2(PCV2) ORF2s gene was induced by IPTG.Inclusion body of the recombinant protein was purified and its reactonogenicity confirmed by Western-blot assay.Purified protein was used as antigen to develop an indirect enzyme-linked immunosorbent assay(ELISA) for detecting serum antibody against PCV2.The indirect ELISA was used to detect 179 serum samples collected from Jilin province and 42.5% samples were positive.The result showed that ...