小鼠仙台病毒ELISA抗体检测规范化试剂盒研制和应用.docxVIP

小鼠仙台病毒ELISA抗体检测规范化试剂盒的研制与应用 向志光，佟巍，刘先菊，张丽芳，李雨函，王艳蓉，刘云波，魏强* * [基金项目]中央级公益性科研院所基本科研业务费专项基金项目（DWS201213） [通讯作者]魏强，Email: weiqiang0430@ （中国医学科学院/北京协和医学院医学实验动物研究所，卫生部实验动物检测中心，北京100021） 【摘要】目的 按照实验动物国家标准和农业部对于兽医诊断制品的要求研制小鼠仙台病毒抗体检测试剂盒，并在临床检测中分析其适用性。方法 建立仙台病毒的种子批和BHK-21细胞的细胞库；标化仙台病毒生产工艺、抗原蛋白纯化工艺；优化ELISA反应板体系；标化质控血清。使用规范化的ELISA试剂盒对我单位672份送检血清样品进行检测，使用IFA和Western blot方法进行复检。结果 病毒的种子批检验表明在-80℃保存半年以上毒力稳定；病毒生产和抗原纯化工艺的标准化提高了抗原生产的稳定性；对照体系的设定降低了环境等变量对于结果判定的影响。在对临床样本的检测过程中发现3种方法的灵敏度ELISA高于Western blot高于IFA。结论 规范化的小鼠仙台病毒ELISA抗体检测试剂盒能对小鼠仙台病毒感染状况作出准确判断，具有一定的稳定性和结果可重复性。 【关键词】小鼠；仙台病毒；ELISA；规范化 【中图分类号】R-332 The development and application of standardized ELISA kit for mouse Sendai virus XIANG Zhiguang, TONG Wei, LIU Xianju, ZHANG Lifang, LI Yuhan, WANG Yanrong, LIU Yunbo, WEI Qiang （Institute of Laboratory Animal Science, Peking Union Medical College/Chinese Academy of Medical Science, Laboratory Animal Monitoring Center, Ministry of Health, Beijing 100021, China） 【Abstract】Objective To develop a standardized ELISA kit for mouse Sendai virus and test it in clinical application. Methods Sendai virus seed batches and BHK-21 cell banks were set firstly; then the crafts for virus production and antigen purification were established; the detection system in ELISA flat was also optimized; the positive and negative serum controls were normalized. A totally 672 sera samples were analyzed by the current ELISA kit, and the results were confirmed by IFA and Western blot. Results the Sendai virus seed batches stored at -80℃ were stable for more than 6 months; the virus production craft and antigen purification craft improved the antigen quality; the control systems reduced the effects from environment. The detection sensitivity of ELISA kit is higher than Western blot, much higher than IFA. Conclusion the standardized mouse Sendai virus detection ELISA kit can detect the Sendai virus infection condition, and the detection result is stable and reproducible. 【Key words】Mouse; Sendai virus; ELISA; St