实验动物科学

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实验大鼠肺炎病毒ELISA检测试剂盒的研究

吴惠英;陈永刚;张然;刘佐民;   

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  • 收稿日期:1992-09-25 修回日期:1992-09-25 出版日期:1992-09-25 发布日期:1992-09-25

Study on Application of ELISA Kit for the Detection of Antibody to PVM in Rat

Wu Huiying Chen Yonggang Zhang Ran Liu Zuomin National Institute for The Control of Pharmaceutical and Biological Products, Beijing 100050   

  • Received:1992-09-25 Revised:1992-09-25 Online:1992-09-25 Published:1992-09-25

摘要: 在小鼠肺炎病毒的抗原特性进行鉴定的基础上,将其毒种感染BHK_(21)细胞作为抗原,建立了检测大鼠PVM抗体的酶联免疫吸附试验的试剂盒。经过提纯PVM免疫血清结合辣根过氧化物酶,用于阻断试验、免疫荧光抗体(IFA)试验,考核了本试剂盒的特异性,对该试剂盒的灵敏性及重演性作了验证。并用本试剂盒对154只大鼠的PVM抗体水平进行调查,结果是开放大鼠PVM阳性率为38.2%,清洁大鼠为0。并对154只大鼠标本用光密度OD值定值的两种方法作了探讨。

关键词: PVM, ELISA阻断试验, OD定值方法

Abstract: Based on the identification of antigenicity of PVM, a ELISA Kit, in which antigen was prepared by BHK21 cells inoculated with PVM, was establishment for detecting antibody to PVM in rats The conjugate (specific antibody labled with HRP) was used in ELISA-block test. IFA test was used for further verification of specificity of ELISA Kit. Antibody protein can be detected at a level of 2μg/ml. The results showed the high sensitivity and good reproducibility. This Kit was used in rountine monitoring. The results indicated that the infection rate was 38.2% (55/114) in conventional rats and 0% (0/10) in clear rats. Compared with two methods for selection of critical value by using OD value from 154 rats no significant diference was found.

Key words: PVM, antibody, ELISA-block, critical value