茨城病病毒S7基因的截短表达及间接ELISA检测方法的建立

Establishment of an indirect ELISA method using a truncated Ibaraki virus VP7 protein as antigen

为建立以茨城病病毒(IBAV)VP7蛋白为诊断抗原的茨城病(IBAD)血清学检测方法,本研究克隆了VP7蛋白抗原性、亲水性较强部分的编码基因,并在原核表达系统中表达了可溶性截短VP7蛋白。用截短VP7蛋白建立了IBAV间接ELISA检测方法。确定的阳性血清的判定标准为不低于0.32。特异性试验表明建立的ELISA方法可以特异性检测IBAV阳性血清。建立的ELISA方法与中和试验结果比较,符合率为77%。用建立的ELISA方法检测70份来自云南的牛血清样品进行检测,IBAV血清阳性率为45.7%。该方法的建立为IBAD的诊断提供了一种简单快速的辅助手段。

The VP7 protein of Ibaraki virus（IBAV） is highly conserved and could be used as diagnostic antigen in serological test.In this study,a truncated VP7 protein gene which retained antigenicity and hydrophilicity of the VP7 protein was cloned and expressed in E.coli.The expressed protein was used to establish an indirect ELISA method.The assay could specifically detect IBAV positive serum,and showed 77 % accordance with neutralization assay.The ELISA method was used to analyze 70 bovine serum samples in Yunnan province and 45.7 % IBAV seropositivity rate was detected.This method could provide an easy and effective approach for the diagnosis of IBAD.