基于快速PCR方法的人参属药用植物鉴别研究  

Application of Rapid PCR Technology on Authentication of Species in Panax Genus

作　　者：崔占虎[1] 袁媛[2] 张景景[3] 王旭[3] 丁生晨 黄显章[3] 蒋超[2] 

机构地区：[1]南阳市第一人民医院,河南南阳473010 [2]道地药材国家重点实验室培育基地/中国中医科学院,北京100700 [3]南阳理工学院,河南南阳473004

出　　处：《中药材》2015年第8期1634-1638,共5页Journal of Chinese Medicinal Materials

基　　金：中医药行业专项(201407003)

摘　　要：目的:建立一种准确、快速、高效鉴别人参属药用植物的分子鉴别方法。方法:采集不同产地的人参、西洋参及同属近缘种植物,所有样品进行总DNA的提取并使用mat K片段进行扩增、测序,进行同源比对后根据其变异位点设计西洋参特异性鉴别引物,人参以文献报道的特异鉴别引物为基础,分别采用两步法进行PCR扩增,从而对人参、西洋参及同属近缘种进行鉴别。结果:通过对影响PCR反应时间的退火温度、变性温度、退火时间、变性时间、循环次数等因素进行优化,并对不同型号PCR仪进行考察,分别获得人参、西洋参快速PCR反应程序。在PCR产物中加入SYBR GreenⅠ染料,正品显示出明亮绿色荧光,而近缘种不显示荧光。结论:快速PCR方法可以简单快速鉴别人参、西洋参,为实现药材分子鉴别的现场运用提供技术支撑。Objective：To establish an accurate, rapid and efficient method for authentication of Panax species by using PCR ampli- fication of specific alleles. Methods ： The samples of Panax species were collected for extracting the total DNA. matK sequence from the Panax species was amplified by PCR and sequenced directionally, and then aligned by using Clustal W. Specific primers were designed and amplified by two-steps PCR amplification method. Results：By optimizing the denatured and annealing temperature and time, cycle numbers ,the rapid PCR methods for authentication of Panax species were established respectively. When SYBR Green I was added in the PCR product, strong green fluorescence was visualized under 365 nm UV lamp whereas adulterants were not. Conclusion ：The rapid PCR method can identify the Panax species rapidly. This study provides the technical support for authentication of Chinese medicinal materials.

关 键 词：快速PCR 人参属 分子鉴定 荧光检测 

分 类 号：R282.5[医药卫生—中药学]