马疱疹病毒1型gG蛋白间接ELISA检测方法的建立及应用

Establishment and Application of an Indirect ELISA Method Using Equine Herpesvirus Type 1 gG Protein

为了解新疆马疱疹病毒1型(EHV-1)的流行情况,需要建立一种检测抗体的间接ELISA方法。采用纯化后EHV-1 XJ2015株的gG蛋白作为检测抗原,优化反应条件后建立检测EHV-1血清抗体的间接ELISA方法,并对该方法进行特异性、敏感性和重复性试验及商品化试剂盒的应用效果对比。结果表明,该方法仅与EHV-1阳性血清发生反应,不与马疱疹病毒4型、马流感病毒和马动脉炎病毒阳性血清发生反应,血清稀释1∶1600后,仍可以检测到阳性,组内及组间变异系数均小于5%。与试剂盒进行检测结果比较,符合率为93.35%。建立了EHV-1 gG蛋白间接ELISA检测方法,可为EHV-1的快速诊断、流行病学调查及防控工作提供技术支撑。

In order to understand the prevalence of Equine herpes virus 1(EHV-1) in Xinjiang, an indirect ELISA method for detecting antibodies should be established urgently.In this study, the purified gG protein of EHV-1 XJ2015 was used as the detection antigen.The reaction conditions, the specificity, sensitivity and repeatability tests of the method were optimized.In addition, the application effect was compared with a commercial kit.The results showed that EHV-1 positive sera can be detected while the positive sera of Equine herpesvirus 4,Equine influenza virus and Equine arteritis virus can’t be detected.The positive sera can still be detected after dilution 1∶1600.The coefficients of variation of inter-group and intra-group are less than 5%.The coincidence rate of the test results of 316 horse sera by this method and a commercial kit is 93.35%.In conclusion, the indirect ELISA detection method used EHV-1 gG protein is established successfully, which will provide technical support for the rapid diagnosis, epidemiological investigation, prevention and control of EHV-1.