PCR方法快速鉴别绞股蓝与乌蔹莓

Rapid PCR identification of Gynostemma pentaphyllum and Cayratia japonica

建立绞股蓝与乌蔹莓2种中药相互鉴别的特异性PCR方法,利用SYBR Green I染料法建立对绞股蓝与乌蔹莓的快速鉴别方法。采集不同产地的绞股蓝与乌蔹莓各9份,通过对其叶绿体DNA中trnL-F片段进行扩增、测序,进行多重序列比对后,根据其变异位点设计相互鉴别的特异性引物,建立特异PCR鉴别方法,并通过加入SYBR Green I染料法建立对2种中药进行快速检测方法,本研究建立了对绞股蓝与乌蔹莓2种中药进行分子鉴别方法,并建立了荧光快速检测方法。

To rapidly and accurately distinguish between the two Chinese medicinal materials, Gynostemma pentaphyllum and Cayratia japonica, a PCR methodology based on the SNP in trnL-F sequence was established. The herbal samples from different production regions were collected for total DNA extraction and trnL-F gene sequencing. The SNPs in trnL-F sequence were determined by using the ClustulX 2.1 program. The primers for the identification were selected by comparing the SNP sites of the herbs to design PCR methods specific to G. pentaphyllum and C. japonica, by addition of SYBR Green I dye to production of PCR, a rapid detection was achieved to positively distinguish the two plant materials.