中药全蝎快速鉴定方法的开发与应用  

Development and Application of Rapid Identification Method for Chinese Medicine Scorpion

作　　者：孟媛 孙淼 吴楠 周童 李洋洋 张宇彤 李明成 高丽君[1] 艾金霞 MENG Yuan;SUN Miao;WU Nan;ZHOU Tong;LI Yang-yang;ZHANG Yu-tong;LI Ming-cheng;GAO Li-jun;AI Jin-xia(School of Medical Technology,Beihua University,Jilin 132013,China;Innovation Center of DNA Fingerprint Detection Technology of Traditional Chinese Medicine,Beihua University,Jilin 132013,China)

机构地区：[1]北华大学医学技术学院,吉林吉林132013 [2]北华大学吉林省中药DNA指纹检测技术科技创新中心,吉林吉林132013

出　　处：《中国药学杂志》2023年第5期396-402,共7页Chinese Pharmaceutical Journal

基　　金：吉林省科技发展计划项目资助(20200404152YY,20200403047SF);北华大学研究生创新计划项目资助(北华研创合字[2021]064)。

摘　　要：目的构建一种能够可视化快速鉴定中药全蝎及其伪混品的方法。方法应用改良十二烷基硫酸钠碱变性(SDS)法提取基因组DNA。选取细胞色素C氧化酶亚基Ⅰ(COⅠ)基因序列设计全蝎特异性引物,进行标记。优化聚合酶链式反应(polymerase chain reaction,PCR)体系,克隆制备全蝎阳性质控品作为对照。构建PCR产物检测试纸条,鉴定市售样品的真伪。结果只有正品中药全蝎在286 bp处扩增出特异性条带,且试纸条的检测结果为阳性,检测灵敏度达到1.0×10^(-2)ng·μL^(-1)。20个全蝎市售样品中18个正品样品,2个不合格样品。结论建立的中药全蝎可视化试纸条检测的方法特异、灵敏、稳定、简单、快速,为中药全蝎的真伪鉴定提供了安全可行的方法。OBJECTIVE To establish a visual and rapid identification method for scorpion and its adulterants.METHODS Genomic DNA was extracted by improved SDS method.The COI gene sequence was selected to design specific primers for scorpion and marked.Optimization of PCR reaction system,cloning preparation of scorpion positive quality control products as con-trol.The PCR product test strip was constructed to identify the authenticity of commercial samples.RESULTS Only the scorpion of the genuine Chinese medicine amplified a specific band at 286 bp,and the detection results of the test strip were positive,with the detection sensitivity of 1.0×10^(-2)ng·μL^(-1).Among the 20 commercial scorpion samples,18 were genuine and 2 were counterfeit.CONCLUSION The established method is specific,sensitive,stable,simple and rapid,which provides a safe and feasible method for the authenticity identification of Chinese medicine scorpion.

关 键 词：全蝎 改良十二烷基硫酸钠碱变性法 聚合酶链式反应 分子克隆 核酸试纸条 

分 类 号：R282[医药卫生—中药学]