Song Yinsen, Yang Zhenzhen, Wang Peipei, Song Ke, Zhang Sisen, Fan Tianli
The Fifth Clinical Medical College of Henan University of Chinese Medicine (Zhengzhou People's Hospital), Translational Medicine Research Center, Zhengzhou, China.
School of Basic Medical Sciences, Zhengzhou University, Zhengzhou, China.
Ann Transl Med. 2022 Jun;10(12):706. doi: 10.21037/atm-22-2415.
(Mu Tong) is commonly misused by (Guan Mutong) and (Chuan Mutong), which are nephrotoxic and carcinogenic. However, in the Pharmacopoeia of the People's Republic of China (2015 Edition), the method for determining remains undefined.
We used DNA barcode-based next-generation sequencing (NGS) combined with quantitative real-time polymerase chain reaction (qPCR) to detect in Longdan Xiegan Wan (LDXGW) for the first time. Compared with chromatographic studies, NGS enables better evaluation of the ingredient components of traditional Chinese medicine (TCM) preparations. The feasibility of qPCR using species-specific primers to determine the authenticity of species has been validated. In this study, the constituents of in LDXGW from three different manufacturers were scanned by NGS. The independently developed qPCR detection primers of , , and were specifically used to analyze the LDXGW mentioned above.
The results showed that qPCR detected in all commercial samples. Meanwhile, NGS detected the counterfeit species (Tie-Xian Lian) in all samples. We found that qPCR shows a difference in detecting , but it was not able to identify the unknown additives and adulterants for the primer pairs of .
Hence, it is sensitive and rapid, qPCR is not suitable for detection alone. The NGS approaches offer important novel insights that complement the qPCR method. The combination of NGS and qPCR will be a powerful complement to traditional identification methods of TCM substances.
（木通）常被肾毒性和致癌性的（关木通）和（川木通）误用。然而，在《中华人民共和国药典》（2015年版）中，测定（木通）的方法仍未明确。
我们首次使用基于DNA条形码的下一代测序（NGS）结合定量实时聚合酶链反应（qPCR）来检测龙胆泻肝丸（LDXGW）中的（木通）。与色谱研究相比，NGS能够更好地评估中药（TCM）制剂的成分。使用物种特异性引物的qPCR测定物种真实性的可行性已得到验证。在本研究中，通过NGS扫描了来自三个不同制造商的LDXGW中（木通）的成分。独立开发的（木通）、（关木通）和（川木通）的qPCR检测引物专门用于分析上述LDXGW。
结果表明，qPCR在所有商业样品中均检测到（木通）。同时，NGS在所有样品中均检测到伪品（铁线莲）。我们发现qPCR在检测（木通）时存在差异，但对于（木通）的引物对，它无法识别未知的添加剂和掺假物。
因此，qPCR灵敏且快速，但不适合单独检测。NGS方法提供了重要的新见解，对qPCR方法起到补充作用。NGS和qPCR的结合将成为中药物质传统鉴定方法的有力补充。
