Jia Jing, Xu Zhichao, Xin Tianyi, Shi Linchun, Song Jingyuan
Key Laboratory of Bioactive Substances and Resources Utilization of Chinese Herbal Medicine, Ministry of Education, Institute of Medicinal Plant Development, Peking Union Medical College and Chinese Academy of Medical SciencesBeijing, China.
Front Plant Sci. 2017 May 31;8:926. doi: 10.3389/fpls.2017.00926. eCollection 2017.
Substandard traditional patent medicines may lead to global safety-related issues. Protecting consumers from the health risks associated with the integrity and authenticity of herbal preparations is of great concern. Of particular concern is quality control for traditional patent medicines. Here, we establish an effective approach for verifying the biological composition of traditional patent medicines based on single-molecule real-time (SMRT) sequencing and DNA barcoding. Yimu Wan (YMW), a classical herbal prescription recorded in the Chinese Pharmacopoeia, was chosen to test the method. Two reference YMW samples were used to establish a standard method for analysis, which was then applied to three different batches of commercial YMW samples. A total of 3703 and 4810 circular-consensus sequencing (CCS) reads from two reference and three commercial YMW samples were mapped to the ITS2 and regions, respectively. Moreover, comparison of intraspecific genetic distances based on SMRT sequencing data with reference data from Sanger sequencing revealed an ITS2 and intergenic spacer that exhibited high intraspecific divergence, with the sites of variation showing significant differences within species. Using the CCS strategy for SMRT sequencing analysis was adequate to guarantee the accuracy of identification. This study demonstrates the application of SMRT sequencing to detect the biological ingredients of herbal preparations. SMRT sequencing provides an affordable way to monitor the legality and safety of traditional patent medicines.
不合格的传统专利药品可能会导致全球范围内与安全相关的问题。保护消费者免受与草药制剂的完整性和真实性相关的健康风险是备受关注的问题。传统专利药品的质量控制尤其令人担忧。在此，我们基于单分子实时（SMRT）测序和DNA条形码技术建立了一种验证传统专利药品生物成分的有效方法。选择《中国药典》中记载的经典草药方剂益母丸（YMW）来测试该方法。使用两个益母丸参考样品建立了一种标准分析方法，然后将其应用于三批不同的市售益母丸样品。分别从两个参考益母丸样品和三个市售益母丸样品中获得的3703条和4810条环形一致序列（CCS）读数被映射到ITS2和 区域。此外，基于SMRT测序数据的种内遗传距离与桑格测序的参考数据比较显示，ITS2和 基因间隔区表现出较高的种内差异，变异位点在物种内显示出显著差异。使用CCS策略进行SMRT测序分析足以保证鉴定的准确性。本研究证明了SMRT测序在检测草药制剂生物成分中的应用。SMRT测序提供了一种经济实惠的方法来监测传统专利药品的合法性和安全性。
