西尼罗河病毒病竞争ELISA检测方法的初步建立及应用

摘要

本研究用表达的西尼罗河病毒（WNV）囊膜蛋白E结构域Ⅲ蛋白作为包被抗原，单抗8F4A4作为竞争检测抗体，初步建立了能够对乙型脑炎病毒（JEV）和WNV进行鉴别诊断的竞争ELISA方法。优化的最佳抗原包被浓度为530ng／mL，单抗的最佳稀释倍数为1：8000，待栓血清的最佳稀释倍数为1：10。通过对56份阴性样品进行检测确定了该方法的临界值为30％，以此为判定标准对临床220份血清进行检测，结果均为阴性。此方法的建立弥补了商品化试剂盒不能区分JEV和WNV感染的缺陷，为我国西尼罗河病毒病的流行病学调查提供了一种有效的抗体检测方法。%A cELISA for detection of WNV antibody was established using purified WNV E protein domain IH as antigen coating the ELISA plate and monoclonal antibody 8F4A4 specific to WNV as competitive detection antibody.The results showed that the antigen coat concentration was 530 ng/mL, the best dilution of McAb 8F4A4 was 1：8000 and the best serum dilution was I：10.A total of 56 clinically confirmed negative control sera were tested by the cELISA.Through analysis, it was concluded that the cut-off value was 30%.A set of 220 clinical serum samples were tested by the cELISA and all were negative.Contrast to the commercial kits, the method could differentiate the JEV and WNV antibody, providing an efficient antibody detection method for the West Nile virus disease epidemiology investigation in our country.