基于LAMP的中药材川贝母真伪鉴别方法的建立  

Authentication of Fritillaria cirrhosa based on loop-mediate isothermal amplification

作　　者：兰青阔 赵新 陈锐 刘征辉 朱珠 王成 沈晓玲 刘娜 王燕 陈利智 王永 LAN Qing-kuo;ZHAO Xin;CHEN Rui;LIU Zheng-hui;ZHU Zhu;WANG Cheng;SHEN Xiao-ling;LIU Na;WANG Yan;CHEN Li-zhi;WANG Yong(Tianjin Institute of Agricultural Quality Standard and Testi ng Technology,Tianjin 300381,China;Tianjin Agricultural University,Tianjin 300384,China)

机构地区：[1]天津市农业质量标准与检测技术研究所,天津300381 [2]天津农学院农学系,天津300384

出　　处：《药物分析杂志》2019年第3期551-556,共6页Chinese Journal of Pharmaceutical Analysis

基　　金：科技部国际合作项目:农产品质量安全分子检测技术研究(2006DFA32380);天津市科技计划项目:中药质量现状评价与标准研究(05SYSYGX25000)

摘　　要：目的:基于荧光-环状等温扩增技术,建立川贝母快速检测方法,实现中药材川贝母的荧光可视化快速鉴定。方法:依据川贝母物种特异性序列设计5套LAMP引物,并测试方法特异性和灵敏度。结果:引物Set1扩增效率最高;使用该引物建立的川贝母LAMP检测方法仅能够在6种川贝母基源药材中扩增出扩增曲线和荧光变化,在伊贝母、浙贝母、平贝母等7种常见混淆品中无扩增产物,方法特异性良好;方法检测灵敏度为0.1%。结论:本研究建立的基于LAMP的川贝母鉴定方法具有较好的特异性和灵敏度,其快速、操作简单、易观察的优点,填补了中药材川贝母真伪快速筛查的技术空白。Objective:To establish rapid visual identification method for Fritillaria cirrhosa based on loop-mediated isothermal amplification. Methods:According to the DNA sequence of the target,five sets of primers were designed and screened by amplification efficiency. Specificity and sensitivity of the method was validated. Results:The LAMP detection method established by using primer set 1 showed best performance on amplification efficiency,specificity and sensitivity. Turbidity and fluorescence changes could be observed in 6 kinds of Fritillariae,and there was no amplification product in 7 common adulterants. The sensitivity of the LAMP method was 0.1%.Conclusion:The proposed LAMP method is specific,sensitive,accurate and simple and easy to observe,which fills the gap for the authentication of Fritillaria cirrhosa.

关 键 词：川贝母 环状等温扩增技术 真伪鉴定 检测方法 分子检测 

分 类 号：R917[医药卫生—药物分析学]