Li Dak Sum Yip Yio Chin R & D Centre for Chinese Medicine, State Key Laboratory of Research on Bioactivities and Clinical Applications of Medicinal Plants, The Chinese University of Hong Kong, Shatin, N.T., Hong Kong, China; School of Life Sciences, The Chinese University of Hong Kong, Shatin, N.T., Hong Kong, China.
Biotechnol Adv. 2019 Dec;37(8):107450. doi: 10.1016/j.biotechadv.2019.107450. Epub 2019 Sep 12.
Conventional Sanger Sequencing for authentication of herbal products is difficult since they are mixture of herbs with fragmented DNA. Next-generation sequencing (NGS) techniques give massive parallelization of sequencing reaction to generate multiple reads with various read length, thus different components in herbal products with fragmented DNA can be identified. NGS is especially suitable for animal derived products with the lack of effective markers for chemical analysis. Currently, second generation sequencing such as Illumina Sequencing and Ion Torrent Sequencing, and third generation sequencing such as PacBio Sequencing and Nanopore Sequencing are representative NGS platforms. The constructed library is first sequenced to obtain a pool of genomic data, followed by bioinformatics analysis and comparison with DNA database. NGS also facilitates the determination of contaminant which is essential for quality control regulation in Good Manufacturing Practice (GMP) factory. In this article, we provide an overview on NGS, summarize the cases on the use of NGS to identify herbal products, discuss the key technological challenges and provide perspectives on future directions for authentication and quality control of herbal products.
传统的桑格测序法很难用于草药产品的鉴定,因为它们是由草药混合物和碎片化 DNA 组成的。新一代测序(NGS)技术可以大规模并行测序反应,生成具有不同读长的多个读段,从而可以鉴定草药产品中具有碎片化 DNA 的不同成分。NGS 特别适用于动物源性产品,因为这些产品缺乏有效的化学分析标记物。目前,Illumina 测序和 Ion Torrent 测序等第二代测序,以及 PacBio 测序和 Nanopore 测序等第三代测序是代表性的 NGS 平台。首先对构建的文库进行测序,以获得一组基因组数据,然后进行生物信息学分析,并与 DNA 数据库进行比较。NGS 还可以方便地确定污染物,这对于良好生产规范(GMP)工厂的质量控制监管至关重要。本文概述了 NGS,总结了 NGS 用于鉴定草药产品的案例,讨论了关键的技术挑战,并对未来草药产品鉴定和质量控制的方向提供了展望。