马疱疹病毒抗体间接ELISA检测方法的建立

Establishment of indirect ELISA for detection of antibodies against equine herpesviruses

为建立一种检测马疱疹病毒抗体间接ELISA方法,采用纯化后的马疱疹病毒1型和马疱疹病毒4型g D蛋白作为抗原,经方阵滴定法优化ELISA反应条件,并对该方法进行特异性和重复性试验。结果显示,使用此方法检测其他马常见病毒病阳性血清均为阴性;组内、组间变异系数均小于10%。采用建立的方法与国外商品化的马疱疹病毒诊断试剂盒对送检的300份马血清进行检测比较,两者符合率为95%。结果表明,本次试验建立的间接ELISA方法可以用来检测马疱疹病毒抗体,为马鼻肺炎的防控工作提供技术支撑。

In order to establish an indirect ELISA method for detection of equine herpesvirus antibody, the glycoprotein D （gD） of equine herpesviruses type 1 （ EHV1 ） and equine herpesviruses type 4 （EHV4） with specific epitope was used as a detection antigen. After optimizing ELISA reaction conditions, an indirect ELISA method was successfully developed. Specificity and reproducibility tests were determined. The results showed that the EHV gD only reacted with antibodies against EHV, but not with antibodies against EIV, EIAV and EAV. The intra-batch and inter-batch variation coefficiencies were lower than 10%. Agreement rate of the indirect ELISA relative to commercial EHV antibody kit was above 95% in detection of 300 horse sera. The results indicated that the indirect ELISA method could be used for detection of the antibody against EHV, which offers a technological support for control of equine rhinopneumonitis.