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中成药大黄蛰虫丸的宏条形码基原鉴定
田晓轩, 刘杰, 窦永杰, 赵亚楠, 吕朝耕, 张春晖, 朱彦
天津中医药大学, 天津市现代中药重点实验室-省部共建国家重点实验室 (培育), 天津 300193
摘要:
[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。
关键词:  中成药  大黄蛰虫丸  宏条形码  分类单元匹配  遗传分析
DOI:10.11656/j.issn.1672-1519.2014.04.14
分类号:
基金项目:国家自然科学基金项目(81202874),高等教育博士学科点专项科研基金(20121210120006),国家科技支撑计划项目(2012BAI29B01)。
The origin identification of Chinese traditional medicine Dahuang Zhechong pill based on metabarcoding method
TIAN Xiao-xuan, LIU Jie, DOU Yong-jie, ZHAO Ya-nan, LV Chao-geng, ZHANG Chun-hui, ZHU Yan
Tianjin State Key Laboratory of Modern Chinese Medicine, Tianjin University of Traditional Chinese Medicine, Tianjin 300193, China
Abstract:
[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.
Key words:  Chinese traditional patent medicine  Dahuang Zhechong pill  metabarcoding  taxonomy assignment  genetic analysis
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