摘要
将盐析法提纯的、用Hela细胞系增殖的猪日本乙型脑炎病毒JapaneseBencephalitis(JEV)抗原包被酶标板,以HRP-SPA作为第二抗体,建立了检测JEV抗体的ELISA试验方法。结果表明,抗原的最适包被浓度为1∶800,待检血清最适稀释度为1∶50,HRP-SPA最佳工作浓度为1∶1000,各步反应时间确定为5min。交叉试验、阻断试验、重复性试验、敏感性试验证明本实验特异性强,重复性好,敏感性高。包被的病毒抗原经甲醛固定15min即可进行正式试验。确定其测定标准为:S/N≥14.81为阳性,S/N<12.53为阴性,12.53≤S/N<14.81为可疑。
The HRPSPA ELISA was established for detecting Japanese B Encephalitis (JEV) antibody.JEV antigen was propagated in Hela cell and purified by salting out method.HRPSPA was used as second antibody.Chess titration test obtained optimal working requirement,such as antigen concentration 1∶800,serum dilution 1∶50,HRPSPA dilution 1∶1 000.It showed that the ELISA had good repeativity,high specifity and high sensitivity.It was successfully applyed for detecting antibody of JEV.
出处
《西北农林科技大学学报（自然科学版）》
CSCD
北大核心
2002年第6期38-42,共5页
Journal of Northwest A&F University(Natural Science Edition)
基金
国家动物诊断液制备中心资助项目(2000-6)
