蝙蝠蛾拟青霉及金水宝胶囊的DNA条形码鉴定  

DNA Barcoding Identification of Paecilomyces hepiali and Jinshuibao Capsules

作　　者：赵小惠 刘霞 钟承赞 杨明 曾乐 向丽[2] 陈士林[2] ZHAO Xiao-hui;LIU Xia;ZHONG Cheng-zan;YANG Ming;ZENG Le;XIANG Li;CHEN Shi-lin(School of Chemistry,Chemical Engineering and Life Sciences,Wuhan University of Technology,Wuhan 430070,China;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;Jiangxi Guoyao Pharmaceutical Co.Ltd.,Nanchang 330052,China)

机构地区：[1]武汉理工大学化学化工与生命科学学院,武汉430070 [2]中国中医科学院中药研究所中药鉴定与安全性评估重点实验室,北京100700 [3]江西国药有限责任公司,南昌330052

出　　处：《中国实验方剂学杂志》2020年第8期156-162,共7页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：国家中药标准化项目(ZYBZH-C-JX-39);国家自然科学基金项目(U1812403-1);国家“重大新药创制”科技重大专项(2018ZX09201009-006-001);中央级公益性科研院所基本科研业务费专项(ZZ13-YQ-102)。

摘　　要：目的:由于传统方法对发酵菌丝体鉴定困难,采用DNA条形码技术对金水宝胶囊原料菌种蝙蝠蛾拟青霉及相关产品进行快速鉴定。方法:采用内转录间隔区(ITS)序列对收集的蝙蝠蛾拟青霉及其易混淆物种共8种168份样品进行条形码鉴定,并基于ITS序列设计蝙蝠蛾拟青霉特异引物对相关产品进行快速鉴定。结果:44份不同来源蝙蝠蛾拟青霉的ITS序列长度均为499 bp,无变异位点。基于ITS序列可对蝙蝠蛾拟青霉及其7种易混伪品进行区别;通过ITS序列设计的蝙蝠蛾拟青霉特异性引物(ITS-BF/ITS-BR)可特异扩增得到长度102 bp的短片段,采用该片段可快速鉴别蝙蝠蛾拟青霉与其他虫草菌,并可区分市场上相关产品。结论:通过ITS序列及特异性引物可以实现蝙蝠蛾拟青霉及其相关产品的快速鉴定,为金水宝胶囊的生产及质量控制提供参考。Objective:Because traditional methods are difficult to identify the fermentation mycelium,DNA barcoding technology was used to quickly identify the raw material strain Paecilomyces hepiali of Jinshuibao capsules and related products.Method:A total of 168 samples of 8 species of P.hepiali and its confusable species were identified by internal transcribed spacer(ITS)sequences,and based on the ITS sequences,P.hepiali specific primers were designed to quickly identify the related products.Result:The length of ITS sequences in 44 P.hepiali samples from different sources was 499 bp and there was no mutation site.It was shown that P.hepiali could be distinguished from 7 confusable species based on ITS sequences.The specific primer(ITS-BF/ITS-BR)of P.hepiali designed by ITS sequences could be amplified to obtain a short fragment of 102 bp in length,which could be used to rapidly identify P.hepiali from other confusable species,and to distinguish relevant products in the market.Conclusion:The rapid identification of P.hepiali and its related products can be achieved through the ITS sequences and specific primers,which provides a reference for the production and quality control of Jinshuibao capsules.

关 键 词：蝙蝠蛾拟青霉 DNA条形码 内转录间隔区(ITS) 特异引物 冬虫夏草 金水宝胶囊 发酵 

分 类 号：R22[医药卫生—中医基础理论] R931[医药卫生—中医学]