酶联免疫吸附试验检测禽脑脊髓炎抗体方法的研究

Study on ELISA method to detect the antibody of AEV

利用鸡胚成纤维细胞 (CEF)繁育禽脑脊髓炎病毒 (AEV) ,取其培养物 ,经差速离心提纯 ,做为酶联包被杭原 ,建立了检测AEV抗体的间接酶联免疫吸附试验 ,初步确定了各种反应条件 :抗原包被浓度 16 7μg/ml,待检血清 1∶10 0稀释 ,酶标抗体工作浓度 1∶5 0 0。以阴性血清OD值平均值加上 2个标准差作为界值 ,当待测样品OD值大于界值 0 2 7时 ,判为阳性 ,否则判为阴性。经交叉试验、阻断试验、重复试验和平行试验 。

The ELISA method to detect the antibody of AEV was established,which purified the AEV bred by CEF and used it as enveloped antigen Conditions of reaction were as follows: consistence of enveloped antigen was 16 7μg/ml, the unknown serum was dilted by 1∶100, consistence of enzyme-linked antibody activated in 1∶500 Using average OD of negative serum plus the two standard deviations as critical value,it was judged positive when the OD of the unknown serum higher than 0 27, otherwise it was negative It was proved that the method was well special and repeatable by cross experiment, interdiction experiment, repeated experiment and parallel experiment