快速PCR法鉴别鱼腥草与百部还魂的方法研究  

Rapid PCR identification of Houttuynia cordata and Gymnotheca chinensis

作　　者：魏艺聪[1] 袁媛[2] 陈建雄[1] 车苏容[1] 赵玉洋 梁一池[1] 

机构地区：[1]福建中医药大学药学院,福建福州350122 [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培育基地,北京100700

出　　处：《中草药》2016年第12期2163-2166,共4页Chinese Traditional and Herbal Drugs

基　　金：中医药行业专项资助(201407003);校管-科研创新平台专项资助(X2015014-平台)

摘　　要：目的建立鉴别鱼腥草Houttuynia cordata与百部还魂Gymnotheca chinensis的特异性PCR方法。方法采集不同产地的鱼腥草与百部还魂样品各8份,所有样品提取总DNA。通过对其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特异鉴别引物,建立特异PCR鉴别方法,并通过加入SYBR Green I染料法对2种中药进行快速检测。另外,构建多重PCR体系,只经一个PCR反应,就能对鱼腥草与百部还魂进行快速分子鉴定。结果所构建特异PCR与多重PCR体系均能产生鱼腥草185 bp的特异鉴别条带,百部还魂389 bp的特异鉴别条带,SYBR Green I染料可进行快速检测方法。结论建立的鱼腥草与百部还魂2种中药的快速PCR鉴别方法简便、可靠。Objective For rapid identification of Houttuynia cordata and Gymnotheca chinensis, the specific PCR for mutual authentication of them was established based on the SNPs in matK sequence. Methods H. cordata and G chinensis samples from different origins were collected, total DNA of all samples was extracted, and the matK gene was seqenced. SNPs in the matK sequences of all the samples were found by ClustulX 2.1 program. Primers for identifying hr. cordata and G. chinens were designed according to the SNP site, and specific PCR method was established to identify them, for rapid detection by addition of SYBR Green I dye. In addition, constructing a multi-PCR reaction system, and then the PCR reaction system was optimized. Results The band special for H. cordata （185 bp） and band special for G chinensis （389 bp） were found using specific PCR reaction and Iliulti-PCR reaction, and SYBR Green I dye can be used for rapid detection. Conclusion The multi-PCR reaction system could be used to identify H. cordata and G. chinensis.

关 键 词：鱼腥草 百部还魂 特异性PCR 多重PCR体系 MATK 

分 类 号：R282.12[医药卫生—中药学]