Wei Yinghua, Ding Sheng, Chen Gangyi, Dong Juan, Du Feng, Huang Xin, Cui Xin, Chen Rong, Tang Zhuo
Natural Products Research Center, Chengdu Institute of Biology, Chinese Academy of Sciences, Chengdu, Beijing, China.
School of Ethnic Medicine, Chengdu University of Traditional Chinese Medicine, Chengdu, China.
Front Plant Sci. 2022 Jul 28;13:948879. doi: 10.3389/fpls.2022.948879. eCollection 2022.
Bulbus Fritillariae (BF) is a kind of herbal medicine widely used in many countries including China, Japan, Korea, and so on. Among the known BF medicinal materials, Bulbus Fritillariae cirrhosae (BF cirrhosae) was reported to have the best curative effect. Due to the limited resources of BF cirrhosae, a lot of adulterants have emerged in the market, impairing the market order, resource development, and above all, clinical efficacy. Here, a novel nucleic acid amplification technique, Recombinase Assisted Loop-mediated isothermal DNA Amplification (RALA), was used to establish a real-time fluorescence isothermal molecular authentication method for five commonly used BF drugs. Moreover, this RALA-based assay can also be developed as a colorimetric detection method for on-site detection. Both real-time fluorescence and visual methods could detect as low as 0.1% genuine targets in the mixed samples. In summary, we report an isothermal detection system for five kinds of BF circulating in the market, providing a new choice for the molecular identification of BF drugs and showing promise in the laboratory testing as well as field identification of other herbal medicines.
川贝母是一种在包括中国、日本、韩国等许多国家广泛使用的草药。在已知的川贝母药材中，川贝母被报道具有最佳疗效。由于川贝母资源有限，市场上出现了许多掺假品，损害了市场秩序、资源开发，尤其是临床疗效。在此，一种新型核酸扩增技术，重组酶辅助环介导等温DNA扩增（RALA），被用于建立五种常用川贝母药材的实时荧光等温分子鉴定方法。此外，这种基于RALA的检测方法还可以开发成色imetric检测方法用于现场检测。实时荧光和可视化方法在混合样品中都能检测到低至0.1%的真品目标。总之，我们报道了一种针对市场上流通的五种川贝母的等温检测系统，为川贝母药材的分子鉴定提供了新的选择，并在其他草药的实验室检测和现场鉴定中显示出前景。
