蛤蚧及其混伪品基于12S rRNA序列的Bar-HRM鉴定研究
投稿时间:2019-03-25     点此下载全文
引用本文:苏燕燕,丁丹丹,马婷玉,向丽.蛤蚧及其混伪品基于12S rRNA序列的Bar-HRM鉴定研究[J].中国现代中药,2019,21(9):1197-1205
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作者中文名作者英文名单位中文名单位英文名E-Mail
苏燕燕 SU Yan-yan 中国中医科学院 中药研究所 中药鉴定与安全性检测评估重点实验室,北京100700 Key Laboratory of Beijing for Identification and Safety Evaluation of Chinese Medicine,Institute of Chinese Materia Medica,China Academy of Chinese Medical Sciences,Beijing 100700,China  
丁丹丹 DING Dan-dan 中国中医科学院 中药研究所 中药鉴定与安全性检测评估重点实验室,北京100700 Key Laboratory of Beijing for Identification and Safety Evaluation of Chinese Medicine,Institute of Chinese Materia Medica,China Academy of Chinese Medical Sciences,Beijing 100700,China  
马婷玉 MA Ting-yu 中国中医科学院 中药研究所 中药鉴定与安全性检测评估重点实验室,北京100700 Key Laboratory of Beijing for Identification and Safety Evaluation of Chinese Medicine,Institute of Chinese Materia Medica,China Academy of Chinese Medical Sciences,Beijing 100700,China  
向丽 XIANG Li 中国中医科学院 中药研究所 中药鉴定与安全性检测评估重点实验室,北京100700 Key Laboratory of Beijing for Identification and Safety Evaluation of Chinese Medicine,Institute of Chinese Materia Medica,China Academy of Chinese Medical Sciences,Beijing 100700,China 向丽,副研究员,研究方向:中药资源和分子鉴定学;Tel:(010)64013996,E-mail:lxiang@icmm.ac.cn 
基金项目:公益性行业科研专项(201507002-4-1-3)
中文摘要:目的:为实现蛤蚧与其常见混伪品的高效鉴别和掺伪检测,并建立蛤蚧中药材的DNA条形码-高分辨率熔解曲线(Bar-HRM)鉴别方法。方法:本研究收集了7个物种48份动物样品和10份蛤蚧粉。基于12S核糖体核糖核酸(12S rRNA)序列对蛤蚧及其6种常见混伪品进行高分辨率熔解曲线(HRM)鉴定研究并测序验证。采用HRM方法进行灵敏度和掺伪检测分析,并对市场蛤蚧粉进行检测验证。结果:蛤蚧及其6种常见混伪品均可通过12S rRNA-HRM曲线分析进行鉴别;可实现纳克级的准确检测,最低掺伪检测限为1%;10份市售蛤蚧粉中6份质量可疑。结论:基于12S rRNA序列的Bar-HRM技术可实现蛤蚧与其常见混伪品的准确鉴别,在掺伪检测中具有独特优势,对市场药材的快速检测及质量评估有一定应用价值。
中文关键词:蛤蚧  12S核糖体核糖核酸基因  高分辨率熔解曲线  鉴别
 
Identification of Gekko gecko and Its Adulterants Using High Resolution Melting Based on 12S rRNA Regions
Abstract:Objective:To develop the Bar-HRM method for identifying Gekko gecko and its adulteration efficiently.Methods:Forty-eight animal samples and ten samples of commercial G.gecko powder were collected from markets.All the samples were identified using HRM analysis combined with DNA barcode technique based on 12S rRNA region.Besides,sensitivity analysis and adulteration detection of HRM were analyzed in this study.Results:It was demonstrated that G.gecko and six common counterfeits could be clearly distinguished through Bar-HRM analysis based on 12S rRNA region.The sensitivity of 12S-HRM in adulteration detection was 1%.Six samples of commercial G.gecko powder were doubtful in quality.Conclusion:The Bar-HRM method based on 12S rRNA region is a powerful tool for the identification of G.gecko and its common adulterants.
keywords:Gekko gecko  12S rRNA  high resolution melting  identification
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