中国中药杂志

2013, v.38(16) 2571-2576

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PCR增强剂对药材DNA分子鉴定的影响
Effect of PCR enhancer in molecular authenticate of Chinese herbal medicine

蒋超;崔占虎;袁媛;黄璐琦;
JIANG Chao1,CUI Zhan-hu2,YUAN Yuan1*,HUANG Lu-qi1(1.National Resource Center for Chinese Materia Medica,China Academy of Chinese Medical Sciences, Beijing 100700,China;2.Baotou Medical College,Baotou 014060,China)

摘要(Abstract):

评价PCR增强剂对中药材DNA分子鉴定的影响,筛选获得适用于中药材DNA分子鉴定的PCR增强剂。文章分别利用CTAB法和碱裂解法提取180个中药材DNA样品,分析在PCR反应体系中使用PCR增强剂对通用引物ITS2,psbA-trnH,rbcL,matK,trnL-trnF片段PCR成功率的影响及实时荧光定量PCR反应体系中使用增强剂对PCR扩增效率的影响。结果表明BSA与PVP组合可以明显增加ITS2,psbA-trnH,rbcL片段的PCR成功率,且PCR增强剂也能消除位点特异性鉴别假阴性结果,提高分子鉴定的准确性,但降低了实时荧光定量PCR的扩增效率。本研究说明BSA和PVP作为PCR增强剂能增加中药材DNA的PCR扩增成功率和分子鉴别的准确性。
To evaluate the effect of PCR enhancer on molecular identification of Chinese herbal medicine,and select the optimal enhancers suitable for traditional Chinese medicine(TCM),genomic DNA from 180 kinds of Chinese herbal medicine was extract by CTAB method and alkaline lysis method,respectively.PCR success rate of five universal fragments(ITS2,psbA-trnH,rbcL,matK,trnL-trnF) was compared in a PCR system with and without enhancer.PCR efficiency of Real-time PCR was also compared in a PCR system with and without enhancer.Results showed that PCR success rate of ITS2,psbA-trnH,rbcL fragment was increased by using PVP and BSA.The PCR efficiency was decreased by PCR enhancer in Real-time PCR system.The results indicate that BSA and PVP as PCR enhancer can dramatically increase PCR success rate and genotyping accuracy in TCM molecular authentication.

关键词(KeyWords): PCR增强剂;中药材;分子鉴定
PCR enhancer;Chinese herbal medicine;molecular authentication

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基金项目(Foundation): 北京市科技专项“道地中药功能基因组北京市重点实验室2012年阶梯计划项目”

作者(Author): 蒋超;崔占虎;袁媛;黄璐琦;
JIANG Chao1,CUI Zhan-hu2,YUAN Yuan1*,HUANG Lu-qi1(1.National Resource Center for Chinese Materia Medica,China Academy of Chinese Medical Sciences, Beijing 100700,China;2.Baotou Medical College,Baotou 014060,China)

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