石杉碱甲快速检测酶联免疫试剂盒的制备  

Preparation of indirect competitive enzyme-linked immunosorbent assay kit for Huperzine A

作　　者：余宇燕[1] 邹艳辉[1] 邱亚利[1] 滕海英[1] 

机构地区：[1]福建中医药大学药学院,福州350122

出　　处：《中国免疫学杂志》2014年第1期89-92,99,共5页Chinese Journal of Immunology

基　　金：国家自然基金项目(81202914);福建省科技厅重点项目(2011Y0035);福建省教育厅项目(JA12171)

摘　　要：目的：制备并鉴定石杉碱甲单克隆抗体,研制石杉碱甲间接竞争酶联免疫试剂盒.方法：以石杉碱甲人工抗原免疫BALB/c小鼠获得单克隆抗体,采用酶联免疫吸附法制备石杉碱甲试剂盒,对试剂盒各项指标进行评价.结果：试剂盒工作范围为5~2 500 ng/ml,检测限为6.242 ng/ml,与多种生物碱均无交叉反应,试剂盒可在4℃或-20℃下稳定贮存半年以上,石杉碱甲片回收率在94.2%~108.6%之间,变异系数在2.3%~4.8%之间.结论：石杉碱甲ELISA试剂盒快速、灵敏、稳定,可用于石杉碱甲的含量测定.Objective： To prepare a kit for the detection of HupA in HupA tablet by indirect competitive enzyme-linked immunosorbent assay （IC-ELISA）. Methods： BALB/c mice were immunized by the conjugation of HupA-BSA, and monoclonal antibodies （McAb） were generated by hydridoma technique. An ELISA for HupA was established successfully. Results： The working range of kit was 5 -2 500 ng/ml. The analytical sensitivity was 6. 242 ng/ml. There were no cross-reactivity with the other alkaloids. The kits could be stored at 4℃ or - 20℃ for half year at least. The recovery were 94.2% -108.6%, RDS 2.3% -4.8%. Conclusion： HupA kits are rapid, sensitive, good stability for the determination of HupA content.

关 键 词：石杉碱甲 单克隆抗体 酶联免疫吸附法 试剂盒 

分 类 号：R392.33[医药卫生—免疫学]