禽腺病毒4型抗体间接ELISA检测方法的建立

摘要

为建立一种快速检测鸡群中禽腺病毒4型(FAdV-4)的血清学方法,本研究利用纯化的FAdV-4中国流行株作为包被抗原,并对各反应条件进行优化,建立了检测FAdV-4血清抗体的间接ELISA方法.FAdV-4抗原与抗新城疫病毒、禽流感病毒、传染性法氏囊病病毒、传染性支气管炎病毒、减蛋综合征病毒阳性血清均无交叉反应,表明该方法具有较好的特异性;批内和批间重复试验的最大变异系数分别为3.735％和4.373％,显示该方法具有较好的稳定性;利用所建立的ELISA方法对40份来自疑似FAdV-4感染病鸡的血清样品进行检测,检测结果与FAdV-4 PCR检测结果符合率为100％.对我国河南、山东、安徽、山西和江苏等不同地区的676份鸡血清样品进行了FAdV-4流行病学调查,结果显示上述省份均存在不同程度的FAdV-4感染,抗体阳性率在69.3 ％～89.1％.表明该方法为FAdV-4流行病学调查和防控提供了一种快速有效的检测方法.%To develop a rapid and sensitive assay for detection of antibody against Chinese fowl adenovirus serotype 4 (FAdV-4),an indirect ELISA for FAdV-4 antibody detection was established with the purified Chinese FAdV-4 variant as the coating antigen.Under the optimized reacting conditions,the indirect ELISA was specific for the detection of FAdV-4 positive serum,and had no cross-reaction with positive sera to NDV,H9/H5 subtype AIV,IBDV,IBV and EDSV.The intra-and inter-assay coefficient of variation (CV) were 3.735％ and 4.373％,respectively.The coincidence rate between the established assay and FAdV-4 etiological detection was 100％.A total of 676 chicken sera collected from chicken farms in Henan,Shandong,Anhui,Shanxi and Jiangsu provinces were tested for the presence of antibodies against FAdV-4.The results showed that there were different degrees of FAdV-4 infection in these provinces,and the positive rate of antibody against FAdV-4 was 69.3％-89.1％.This study provided an useful tool for FAdV-4 epidemiological investigation and prevention.