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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 To establish a specific PCR method for identifying Houttuynia cordata and Gymnotheca chinensis. Methods Eight samples of Houttuynia cordata Thunb. And 100 Reiki samples from different areas were collected, and the total DNA was extracted from all the samples. The mat K fragments were amplified and sequenced. After homologous comparison, specific primers were designed according to their variation sites, and a specific PCR method was established to detect the two KCRs by SYBR Green I dye method. In addition, the construction of multiplex PCR system, only by a PCR reaction, will be able to Houttuynia and one hundred reincarnation rapid molecular identification. Results The PCR-specific PCR and multiplex PCR systems were able to generate 185 bp Houttuynia cordatae and 389 bp Trionyx spicifaciens respectively. SYBR Green I dye could be used for rapid detection. Conclusion The rapid PCR identification method of two traditional Chinese medicines, Houttuynia cordata Thunb and Resurrection, is simple and reliable.