犬副流感病毒HN基因在昆虫细胞中的表达及其间接ELISA检测方法的建立

Expression of HN protein of canine parainfluenza virus in insect cells and establishment of an indirect ELISA for the antibody detection

为建立检测犬副流感病毒(CPIV)抗体的方法,本研究利用杆状病毒表达系统表达CPIV的HN蛋白,以其为包被抗原,采用方阵法确定包被抗原和血清最佳工作浓度,对各种反应条件进行优化,建立了CPIV血清抗体的间接ELISA检测方法。该方法仅与CPIV阳性血清发生特异性反应,与犬细小病毒(CPV)和犬腺病毒(CAV)的阳性血清无交叉反应;批内和批间变异系数小于10%。利用建立的ELISA方法与血凝抑制试验分别对48份送检犬血清样品进行检测,符合率为92%。本研究为建立快速、准确、简便的犬副流感鉴别诊断试剂盒奠定了基础。

To develop a rapid method for detecting antibody against canine parainfluenza virus （CPIV）, an ELISA was established with the recombinant HN protein of CPIV expressed and purified from baculovirus expression system as coating antigen. The ELISA method was only able to detect antibodies against CPIV specifically, and no cross-reaction was shown with positive sera against canine parvovirus and canine adenovirus. In addition, the coefficients of variation for both inter-assay and intra-assay was less than 10%. Moreover, forty eight clinical sera samples tested by the ELISA and hemagglutination inhibition, the coincidence rate of two methods was 92 %. In conclusion, the ELISA with the recombinant HN as coating antigen was an efficient and sensitive method for serological surveillance of CPIV infection.