摘要
目的:蜈蚣是我国传统的中药材,具有良好的药用价值。目前市场上蜈蚣的混伪品日益增多,临床用药的安全性和有效性难以保证。该研究建立了一种蜈蚣基原物种的特异性聚合酶链式反应(PCR)方法,以准确、简便的鉴别蜈蚣及其常见混伪品。方法:通过比较蜈蚣及其混伪品基原物种的COI基因序列差异,根据变异位点设计蜈蚣正品少棘巨蜈蚣的特异性鉴别引物WG-500.F和WG-500.R,采集蜈蚣原动物样本及药材样本,优化反应体系并对此方法进行耐受性和适用性的考察和验证。结果:少棘巨蜈蚣原动物样本及蜈蚣正品药材使用设计的蜈蚣鉴别引物经PCR扩增和凝胶电泳后出约500 bp的单一明亮条带,其他混伪品如多棘蜈蚣、模棘蜈蚣、哈氏蜈蚣、海南蜈蚣等均无条带出现。结论:PCR鉴别方法可准确鉴别蜈蚣原动物和药材的真伪,具有高度特异性,为中药材蜈蚣的真伪鉴别提供了良好的科学依据。该方法具有简单、直观等特点,有利于广泛的普及与应用,在中药材鉴定方面有着广阔的应用前景。
Objective:Scolopendra was a traditional Chinese medicine(TCM)with a good medicinal value.Nowadays,there have been increasingly more adulterates of Scolopendra in the medicine market.To ensure the safe and effectiveness of clinical medicines,a convenient and accurate specific polymerase chain reaction(PCR)method for identification of medicinal Scolopendra from its common adulterates was established.Method:Based on the differences of COI gene DNA sequences among Scolopendra subspinipes mutilans and adulterants,the specific primer was designed,the reaction conditions were optimized,and the PCR method for identification was explored and verified in terms of tolerance and feasibility.Besides,the original animal samples and medicine of Scolopendra were collected.Result:Through the established PCR reaction system,the bright and simple fragments of 500 bp was amplified from DNA templates of S.subspinipes mutilans.All of the adulterants were negative by the multiplex PCR assay,such as S.multidens,S.subspinipes,S.dehaani,S.hainanum.Conclusion:The identified primer is highly specific,and the specific PCR method established in this paper can accurately identify Scolopendra and its adulterants,so as to provide an excellent scientific basis for the identification of TCM Scolopendra.The method is simple and intuitive,and facilitates wide promotion and application of the method,with a broad application prospect in the identification of TCM.
作者
田娜
袁媛
金艳
杨全
赵玉洋
蒋超
TIAN Na;YUAN Yuan;JIN Yan;YANG Quan;ZHAO Yu-yang;JIANG Chao(School of Traditional Chinese Medicine,Guangdong Pharmaceutical University,Guangzhou 510006,China;State Key Laboratory Breeding Base of Dao-di Herbs,National Resource Center for Chinese Materia Medica,China Academy of Chinese Medical Sciences,Beijing 100700,China)
出处
《中国实验方剂学杂志》
CAS
CSCD
北大核心
2019年第17期113-117,共5页
Chinese Journal of Experimental Traditional Medical Formulae
基金
中央本级重大增减支项目(2060302)
中央级公益性科研院所基本科研业务费专项(ZZ10-008)
关键词
蜈蚣
聚合酶链式反应
少棘巨蜈蚣
混伪品
Scolopendra
polymerase chain reaction(PCR)
Scolopendra subspinipes mutilans
adulterants
作者简介
第一作者:田娜,硕士,从事中药分子鉴定,E-mail:2509769269@qq.com;通讯作者:蒋超,博士,助理研究员,从事中药分子鉴定研究,Tel:010-64087649,E-mail:jiangchao0411@163.com
