【摘要】 为建立一种能够快速鉴别检测猪伪狂犬病毒(PRV)疫苗免疫抗体和野毒抗体的方法，分别对PRV的gD蛋白和gE蛋白进行原核表达，以其重组蛋白作为检测抗原分别建立了gD-间接ELISA方法和gE-间接ELISA方法，并联合应用两种检测方法对临床血清样品进行检测。结果表明：获得了以包涵体形式表达的重组gD蛋白和重组gE蛋白，经Western blot检测表明两种重组蛋白均具有良好的免疫学活性；gD-间接ELISA方法和gE-间接ELISA方法检测PRV阳性血清的最低检出效价分别为1:25 600和1:12 800,gD-间接ELISA方法检测PRRSV、CSFV、PPV、PCV-2、PEDV、TGEV阳性血清均为阴性；gE-间接ELISA方法检测PRV疫苗免疫血清以及PRRSV、CSFV、PPV、PCV-2、PEDV、TGEV阳性血清均为阴性；建立的gD和gE蛋白的两种间接ELISA方法的批内和批间重复性试验的变异系数分别小于5%和7%;联合应用gD-间接ELISA方法和gE-间接ELISA方法检测2156份临床血清样品，PRV抗体阳性率为88.96%,PRV野毒抗体阳性率为4.04%,PRV免疫抗体阳性率为84.93%。说明建立的gD-间接ELISA方法和gE-间接ELISA方法均具有良好的敏感性、特异性、重复性，两种方法的联合应用实现了PRV免疫抗体和野毒抗体的鉴别检测。更多还原

【Abstract】 In order to establish a rapid method for the identification and detection of porcine pseudorabies virus(PRV) vaccine immune antibody and wild virus antibody, the prokaryotic expression technology was used to express the gD and gE protein of PRV respectively. The recombinant gD and gE protein were used as detection antigens, respectively, and the gD and gE indirect ELISA method were established respectively. The clinical serum samples were detected by two detection methods. The results showed that the recombinant gD and gE proteins expressed in the form of inclusion bodies were obtained. The Western blot showed that the recombinant gD and gE proteins had good immunological activity, the lowest detection titers of gD and gE indirect ELISA for PRV positive serum were 1:25 600 and 1:12 800, respectively. The results showed that the positive sera of PRRSV, CSFV, PPV, PCV-2, PEDV and TGEV detected by gD indirect ELISA were all negative; those of PRRSV, CSFV, PPV, PCV-2, PEDV and TGEV positive sera detected by gE indirect ELISA were all negative; the coefficient of variation of intra and inter assay repeatability tests of gD indirect ELISA and gE indirect ELISA were less than 5% and 7%, respectively. The 2156 clinical serum samples were detected by gD and gE indirect ELISA. The positive rate of PRV antibody was 88.96%, the positive rate of PRV wild virus antibody was 4.04%, and the positive rate of PRV immune antibody was 84.93%. The results showed that both gD indirect ELISA and gE indirect ELISA have good sensitivity, specificity and repeatability. The combined application of the two methods realized the differential detection of PRV immune antibody and wild virus antibody.更多还原