| 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Japanese B encephalitis(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∶1 000,各步反应时间确定为5 min.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15 min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. |
| 关键词: 猪 HRP-SPA ELISA JEV抗体 兽医诊断 |
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| 基金项目:国家动物诊断液制备中心资助项目(2000-6) |
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| Research of using HRP-SPA ELISA to detect Japanese B encephalitis antibody |
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| Abstract: |
| The HRP-SPA ELISA was established for detecting Japanese B Encephalitis (JEV) antibody.JEV antigen was propagated in Hela cell and purified by salting out method.HRP-SPA was used as second antibody.Chess titration test obtained optimal working requirement,such as antigen concentration 1∶800,serum dilution 1∶50,HRP-SPA 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. |
| Key words: swine HRP-SPA ELISA JEV antibody veterinary diagosis |
