中成药大黄蛰虫丸的宏条形码基原鉴定

目的]探索应用宏条形码方法进行复方中成药基原鉴定分析的可行性。方法]磁珠法提取中药大黄蛰虫丸DNA,以16S、trnL及Mini-barcode为靶标进行荧光定量PCR。产物经克隆测序拼接后,经BLASTn比对,用MEGAN软件进行分类单元解析,对虻虫来源序列应用MEGA软件进行遗传分析。结果]从23条单克隆测序结果中解析到蛴螬、虻虫、甘草及桃仁或苦杏仁等中药成分,对虻虫6条序列分析,显示其源于3组遗传差异较显著的个体。结论]宏条形码技术可用于中成药成分的鉴定,可作为中成药质量评价的潜在工具。

The origin identification of Chinese traditional medicine Dahuang Zhechong pill based on metabarcoding method

Objective] To investigate the ability of metabarcoding technology as one kind of identification tool for the origin of Chinese traditional patent medicine. Methods] The DNA of TCMDahuang Zhechong pill was extracted by paramagnetic particle method, and the 16S, trnL and Mini-barcode fragments were amplified by real-time PCR. The products of PCR were cloned and sequenced. After BLASTn alignment, the sequences were parsed into taxonomic term by MEGAN. The sequences representing TCM Mengchong were analysed by MEGA. Results] The TCMQicao, Mengchong, licorice and Peach Seed & Bitter Apricot Seed were identified from 23 sequences. Three groups of individuals with significant genetic differences were identified from 6 sequences that representing TCM Mengchong. Conclusion] Metabarcoding technology could be used for component identification of Chinese traditional patent medicine and should be an useful tool for quality evaluation of Chinese traditional patent medicine.