新城疫病毒抗体ELISA检测方法的建立与应用

Establishment and Application of an Indirect ELISA for Detecting Antibodies Against Newcastle Disease Virus

为建立一种快速的新城疫病毒抗体检测方法,本研究以原核表达的重组HN蛋白作为诊断抗原,建立了检测新城疫病毒抗体的间接ELISA诊断方法。该方法检测AIV(H9亚型)、IBV、IBDV、EDSV 4种常见禽病病原的阳性血清均为阴性;检测灵敏度为1∶12 800;批内重复性试验、批间重复性试验的变异系数分别小于5%和10%;与血凝抑制试验(HI)符合率为96.1%。本研究建立的NDV HN间接ELISA检测方法具有良好的特异性、敏感性和重复性,为NDV的抗体检测及流行病学调查等快速诊断提供一种技术手段。

To develop a method to detect the antibody against newcastle disease virus, an indirect ELISA was successfully developed to detect anti-NDV antibody using recombinant HN protein as coating antigen. The method detected the positive sera of other four avian diseases（H9 AIV, IBV, IBDV, EDSV）were negative. Coefficients of variability percent （C V%） of intro-batch and inter-batch duplicative tests were less than 5 % and 10 %, respectively. Lowest antibody titer of positive control serum was 1 ： 12 800. Compared with the HI, the concordance was 96.1%. Therefore, the HN indirect ELISA has good specificity, sensitivity and repeatability, could be used as a simple and rapid serology detection method for monitoring of anti-NDV antibody and epidemiologic survey of NDV.