霍山石斛的PCR-RFLP鉴别研究  

Molecular identification of Dendrobiumhuoshanense by PCR-RFLP analysis

作　　者：胡冲 张亚中 袁媛[2] 蒋超[2] 魏峰[3] 马双成[3] HU Chong;ZHANG Ya-zhong;YUAN Yuan;JIANG Chao;WEI Feng;MA Shuang-cheng(Anhui Institute for Food and Drug Control,Hefei 230051,China;National Resource Center for Chinese Materia Medica,China Academy of Chinese Medical Sciences,Beijing 100700,China;National Institute for Food and Drug Control,Hefei 230051,China)

机构地区：[1]安徽省食品药品检验研究院,合肥230051 [2]中国中医科学院中药资源中心,道地药材国家重点实验室,北京100700 [3]中国食品药品检定研究院,北京100050

出　　处：《药物分析杂志》2020年第12期2109-2115,共7页Chinese Journal of Pharmaceutical Analysis

基　　金：2015年安徽省科技攻关计划项目资助(1501041174)。

摘　　要：目的:建立一种霍山石斛特异性的分子鉴别方法。方法:采用RAD测序技术对霍山石斛及其近伪品进行测序分析,根据差异片段设计聚合酶链反应-限制性内切酶酶切长度多态性(PCR-RFLP)引物,使用限制性内切酶进行酶切鉴别。结果:霍山石斛鉴别引物可扩增霍山石斛及其近伪品序列,扩增产物为153bp大小的条带,而限制性内切酶Alu I可以识别霍山石斛及其伪品的差异序列,仅近伪品石斛的序列可被酶切形成两个片段,而霍山石斛的序列不能被切开,从而特异性鉴别是否为霍山石斛。结论:本实验建立的PCR-RFLP方法可以鉴别霍山石斛的真伪。Objective:To establish a specific DNA molecular identification method for dendrobiumhuoshanense.Methods:Dendrobiumhuoshanense and its adulterants were analyzed by RAD sequencing technology. PCRRFLP primers were designed based on differential fragment,and restriction enzyme digest was further conducted to identify dendrobiumhuoshanense and its adulterants. Results:153 bp specific fragment bands from dendrobiumhuoshanense and its adulterants were amplified by identification primers. AluI restriction endonuclease could only digest the amplified fragment of dendrobiumhuoshanense adulterants in to two smaller pieces and could not digest the amplified fragment of dendrobiumhuoshanense. In this way,dendrobiumhuoshanense was identified specifically.Conclusion:The method of PCR-RFLP developed in this research is an accurate identification method for dendrobiumhuoshanense and its adulterants.

关 键 词：霍山石斛 混伪品 简化基因组测序 聚合酶链反应-限制性内切酶酶切长度多态性 分子鉴定 

分 类 号：R917[医药卫生—药物分析学]