新孢子虫dNcSRS2重组蛋白间接ELISA的建立及其应用

Establishment of Recombinant dNcSRS2 Protein Based Indirect ELISA for Detection of Antibody against Neospora caninum and Its Application

利用新孢子虫体外重组表面蛋白dNcSRS2蛋白作为包被抗原,对各项条件进行优化,确定判定标准,建立了检测新孢子虫血清抗体的间接ELISA方法。经对多例血清检测表明,所建立的诊断试剂盒重复性好、特异性强、灵敏度高,与进口的IFAT及两种商品化ELISA试剂盒的检测结果相比较,符合率均达到92%以上。应用建立的ELISA方法对236份奶牛血清的新孢子虫抗体进行检测,阳性率为22%。这是国内首次利用重组蛋白建立的诊断试剂盒,该方法的建立将为牛新孢子虫病的诊断与流行病学调查提供有效的技术手段。

The purified recombinant dNcSRS2 protein was used as coating antigen to establish an indirect ELISA for detecting antibody against Neospora caninum. Commercially available diagnostic kits （IFAT and two commercial ELISA kits） for N. caninum were chosed as the reference standard methods. The results showed that the ELISA method was highly sensitive, specific and reproducible. The agreements between the methods were above 92%. Dairy cattle sera （n=236） were tested for antibodies to N. caninum; out of which 22 % reacted positively. This is the first use of recombinant protein establishing diagnostic method in China. The new ELISA using dNcSRS2 could be used for mass screening of the prevalence of N. caninum infection in diary cattle.