Real-time fluorescence and colorimetric identification of bulbus fritillariae using recombinase assisted loop-mediated isothermal DNA amplification (RALA).
Real-time fluorescence and colorimetric identification of bulbus fritillariae using recombinase assisted loop-mediated isothermal DNA amplification (RALA).
复制标题
使用重组酶辅助环介导等温 DNA 扩增 (RALA) 实时荧光和比色鉴定贝母
DOI：
10.3389/fpls.2022.948879
复制
发表时间：
2022
影响因子：
5.6
通讯作者：
Tang, Zhuo
中科院分区：
文献类型：
--
作者：
Wei, Yinghua;Ding, Sheng;Chen, Gangyi;Dong, Juan;Du, Feng;Huang, Xin;Cui, Xin;Chen, Rong;Tang, Zhuo
关键词：
贝母（BF）是一种在中国、日本、韩国等国家广泛使用的草药。在已知的BF药材中，据报道川贝母(BF cirrhosae)的疗效最好。由于BF肝硬化资源有限，市场上出现了大量掺假品，影响了市场秩序、资源开发，更影响了临床疗效。在此，采用一种新型核酸扩增技术——重组酶辅助环介导等温DNA扩增（RALA），建立了五种常用BF药物的实时荧光等温分子鉴定方法。此外，这种基于 RALA 的测定还可以开发为用于现场检测的比色检测方法。实时荧光和视觉方法都可以检测到混合样品中低至 0.1% 的真实目标。总之，我们报道了市场上流通的五种BF的等温检测系统，为BF药物的分子鉴定提供了新的选择，并在实验室检测以及其他草药的现场鉴定中显示出应用前景。基于 RALA 的贝母识别图解摘要工作流程。
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. Graphical Abstract Workflow of RALA-based bulbus fritillariae identification.
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