鼠疫耶尔森菌rV抗原单抗系统及其ELISA检测方法的建立  

Development of monoclonal antibodies and ELISA in detection of Yersinia pestis rV antigen

作　　者：常娅莉[1] 焦磊[1] 魏东[2] 吴智远[1] 胡丽娜[1] 卜培英[1] 白钰[3] 王秉翔[1] 

机构地区：[1]兰州生物制品研究所有限责任公司,兰州730046 [2]中国食品药品检定研究院,北京100050 [3]甘肃农业大学生命科学院,兰州730070

出　　处：《微生物学免疫学进展》2012年第3期13-18,共6页Progress In Microbiology and Immunology

基　　金：国家"863"课题(2006AA02Z461)

摘　　要：目的建立ELISA双抗体夹心法,测定重组毒力因子rV抗原含量。方法采用杂交瘤技术,制备鼠疫菌rV抗原的鼠单克隆抗体,对抗原表位和单抗特异性进行分析及鉴定,建立ELISA双抗体夹心法,并验证方法的专属性、准确性、精密度和线性范围。结果成功组建了鼠疫菌rV抗原诊断试剂,灵敏度最低检测值为10 ng/mL。结论该方法可用于免疫学检测鼠疫组分疫苗原液rV抗原含量及制备过程中抗原活性,是鼠疫组分疫苗制备中一种重要的质量控制手段,也为进一步开发鼠疫诊断试剂盒及其他相关研究奠定了基础。Objective To establish a sandwich ELISA in detection of the concentration of rV antigen of Yersinia pestis. Methods The McAbs against rV antigen were prepared by the hybridoma technology and Sandwich ELISA was developed on the basis of the epitope analysis and McAb specificity, in which specificity, accuracy, precision and linear range were v^idated. Results The diagnostic kit for Yersinia pestis rV antigen was successfully set up. The minimum detection con- centration of rV antigen is 10 ng/mL. Conclusion The kit is used in detection of the concentration of rV antigen in stock solution of plague component vaccine and in monitoring of antigeneicity in the preparing process. It is a important method for quality control in preparation of plague component vaccine, this method also provides a foundation for a further develo- ping the plague diagnostic reagents and other related investigations.

关 键 词：鼠疫耶尔森菌 rV抗原 单克隆抗体 表位 双抗体夹心ELISA 

分 类 号：R392.1[医药卫生—免疫学]