猪流行性腹泻病毒血清IgG间接ELISA检测方法的建立

Establishment of an indirect ELISA for detecting serum IgG to porcine epidemic diarrhea virus

以大肠杆菌表达系统制备的猪流行性腹泻病毒(porcine epidemic diarrhea virus,PEDV)中和抗原S1D(1μg/mL)每孔100μL进行包被,利用HRP-兔抗猪IgG(1∶20000)建立了检测猪血清中抗PEDV IgG的间接ELISA方法。阴性判断临界值(M+2SD)为0.632,批内和批间检测的平均变异系数分别为2.3%和3.1%,该方法与商品化的PEDV抗体间接ELISA检测试剂盒同时检测50份临床血清样品,均为阳性,上述结果表明该方法可用于PEDV血清IgG的临床检测。

E.coli-expressed porcine epidemic diarrhea virus(PEDV)neutralization antigen S1D(1μg/mL,100μL per well)and rabbit anti-pig IgG-HRP(1∶20000)were used to establish an indirect ELISA for the detection of anti-PEDV IgG in pig serum.OD450=0.632 was set as the negative cut-off value(M+2SD),and the variation coefficients of intra-and inter batch were 2.3% and 3.1%,respectively.Fifty clinical sera samples were tested with the developed indirect ELISA using approved-commercial PEDV indirect ELISA kit as comparison and all samples were judged positive with each method.Taken together,these results indicated that the indirect ELISA in this study could be used for detecting antibodies to neutralizing antigen PEDV S1D.