应用等位基因特异聚核酶链式反应鉴别半夏类药材

 目的建立能区分半夏正、伪品的分子鉴定方法。方法下载GenBank中半夏属、犁头尖属及天南星属相关序列,利用DNAMAN进行比对分析,根据ITS序列中半夏的SNP位点设计引物PtITS178F和PtITS555R,采用等位基因特异PCR进行扩增。结果对半夏5个产地的新鲜样品、10个产地的药材以及5批虎掌、水半夏样品进行PCR检测。引物PtITS178F和PtITS555R只对半夏类药材扩增395 bp片段,虎掌和水半夏均无扩增条带。结论等位基因特异PCR能准确检测半夏的SNP位点,达到准确鉴别半夏正伪品的目的,是一种有前途的中药材分子鉴定方法。

Authentication of Pinellia ternata and Their Adulterants by AS-PCR

OBJECTIVE To establish a molecular method to authenticate Pinellia ternata and their adulterants. METHODS Download DNA sequences of Pinellia tenore, Typhonium schott and Arisaema martius from GenBank, using DNAMAN to align the sequences. Design allele-specific primers for P. ternata according to its SNPs in ITS sequence. Using primer PtITS178F and PtITS555R to amplify all samples. RESULTS Template DNA of P. ternata containing 5 fresh samples, 10 dried samples from different places could be amplified 395 bp band whereas P. pedatisecta and Typhoniumflagelliforme have no band. CONCLUSION AS-PCR has the advantages of highly specific and good reproducibility, by which P. ternata can be identified from its adulterants quickly. It is a potential method to use in the molecular identification of other meteria medica.