特异性可卡因核酸适体测试条的研制  

Study of aptamer-based immunochromatographic test for detection of cocaine

作　　者：曾立波[1] 胡小龙[1] 张玉荣[1] 丁国荣[1] 张润生[1] 朱凯[1] 梁晨[1] 曹芳琦[1] 

机构地区：[1]上海市公安局物证鉴定中心上海市现场物证重点实验室,上海200083

出　　处：《中国法医学杂志》2014年第5期424-426,共3页Chinese Journal of Forensic Medicine

摘　　要：目的筛选识别可卡因的特异性核酸适体,制备可卡因核酸适体测试条(盒)。方法通过SELEX技术筛选可卡因核酸适体,制备胶体金标记可卡因完全抗原(COC-BSA),制作可卡因层析测试条,并对方法特异性、灵敏度、准确性等性能指标进行考察和验证。结果甲基苯丙胺等64种药品和毒品采用本文研制的可卡因测试条检测,结果均为阴性,无交叉反应;检测可卡因最低浓度在5ng/mL,比胶体金单克隆抗体方法检测高5倍;将测试条密封45℃烘烤1个月后,检测结果与未烘烤过的测试条检测结果一致;与GC/MS检测结果对照,准确性为100%。结论本文研制的可卡因核酸适体检测试条,性能稳定,特异性良好,灵敏性和准确性均可达到实际检测的要求,在毒品、毒物的检测中具有广阔的应用前景。Objective To screen the aptamers specific recognize cocaine and develop a aptamer-based assay for cocaine detection. Methods The aptamers recognize cocaine have been got by SELEX technology. The complete antigen cocaine-BSA was conjugated to colloidal gold particles then used for making the immunochromatographie test strip. The specificity, sensitivity and accuracy for the assay have been evaluated and validated. Results Cocaine aptamer assay can specifically detect cocaine without any cross reactions to 64 kinds of drugs such as methamphetamine. The detection limit for cocaine detection is 5ng/mL which is 5 times more sensitive than the antibody-based assay. The stability experiment showed the aptamer test can maintain its effect for at least one month at 45~C. The test results agreed well with the GC-MS analysis. The accuracy is 100%. Conclusion The aptamer-based cocaine immunoehromatographic test is a stable and specific assay for deIecting cocaine which meets the requirements for practical applications in its sensitivity and accuracy. There will be a good prospect for this assay used for the drugs and toxicant detection.

关 键 词：法医毒物分析 核酸适体 可卡因 胶体金 单克隆抗体 

分 类 号：D919[医药卫生—法医学]