鼠疫抗原单克隆抗体的制备及效果评估

Research on preparation and evaluation of anti-yersinia pestis monoclonal antibody

目的 制备鼠疫F1抗原检测试剂。 方法 用杂交瘤细胞融合技术制备抗鼠疫McAb ;用ELISA法检测其特异性、敏感性 ;用双抗体夹心ELISA法检测鼠疫F1抗原。 结果 抗鼠疫F1抗原McAb效价达 1:1× 10 6,与 9种细菌抗原无交叉反应 ,ELISA法可检测到的鼠疫F1抗原的最小浓度为 0 0 0 65 μg/ml ,经多次验证 ,结果稳定。 结论 该McAb具有高特异性和敏感度 ,所制备的试剂盒有望成为鼠疫流行病学监测和免疫学诊断的重要手段。

Objective To make a diagnostic reagent for the detection of yersinia pestis antigen with anti-yersinia pestis monoclonal antibody(McAb). Methods The hybridoma cell clone secreting anti-yersinia pestis antibody was screened and cloned by using hybridoma cell fusion technique. The specificity and sensitivity of the McAb were identified by ELISA. Sandwich-ELISA was conducted to detect yersinia pestis F 1 antigen. Results The McAb’S work fiter could reach to i:106.And it had no crossing reaction with 9 kinds of bacteria antigen detected by ELISA. The lowest density of yersinia pestis F 1 antigen that can be detected is 0.0065～tg/ml. All the results above were proved to be steady Conclusion The yersinia pestis diagnostic reagent with high specificity and sensitivity is proved probably to be an important method of clinical diagnosis and epidemiologic monitoring of plague.