为建立一种H3N2亚型犬流感病毒(CIV)血清学检测方法,本研究利用CIV重组HA1蛋白作为检测抗原,建立H3N2亚型CIV抗体的间接EHSA检测方法,并优化反应条件。通过检测阴性血清样本30份确定其临界值为0.228。该方法与抗猪流感病毒H1N1、H1N2、H6N6、H9N2的阳性血清和犬瘟热病、犬细小病、犬副流感的阳性血清均无交叉反应。变异系数在1.01%~7.52%之间,具有较好的重复性。通过对150份临床样品进行检测并与血凝抑制试验检测结果比较,总符合率为98.6%。本研究为CIV流行病学调查提供了一种快速、方便、敏感的抗体检测方法。
In this study, an indirect ELISA method for detection of Canine influenza virus (CIV) was developed by using purified recombinant HA1 protein of the virus as coating antigen. It had no cross reaction with positive serum of H1, H6, H9 subtypes of swine influenza virus, and canine distemper virus, canine parvovirus, canine parainfluenza virus. The coefficient variation was between 1.01 and 7.52. A total of 150 clinical samples was detected by the established protocol and compared with hemagglutination inhibition (HI) test. The coincidence rate was 98.6%. The results showed that this indirect ELISA was specific and sensitive for CIV antibody, which would provide a convenient method in studies on the epidemiology and surveillance of canine influenza.