[关键词]
[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matK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。
[Key word]
[Abstract]
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 H. 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 multi-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.
[中图分类号]
[基金项目]
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)