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基于直接环介导等温扩增法的半夏及伪品水半夏的快速比色和荧光鉴定

Rapid colorimetric and fluorescence identification of Pinelliae Rhizoma and adulterate Rhizoma Typhonii Flagelliformis using direct-LAMP assay.

作者信息

Li Huilin, Cui Jiaqi, Chen Hongling, Li Hongying, Xie Yuchen, Song Wenjun, Chen Rong

机构信息

State Key Laboratory of Southwestern Chinese Medicine Resources, Chengdu University of Traditional Chinese Medicine, Chengdu, China.

School of Ethnic Medicine, Chengdu University of Traditional Chinese Medicine, Chengdu, China.

出版信息

Food Chem. 2023 Oct 26;437(Pt 1):137840. doi: 10.1016/j.foodchem.2023.137840.

Abstract

Pinelliae Rhizoma, renowned for its medicinal benefits against cough, insomnia and inflammation, is frequently utilized in botanical dietary supplements and herbal products. However, due to high demand, there is a risk of adulteration. This study introduced Direct-Loop-Mediated Isothermal Amplification (Direct-LAMP), a rapid colorimetric technique, for distinguishing Pinelliae Rhizoma from its primary adulterant, Rhizoma Typhonii Flagelliformis. This method comprises three parts: DNA extraction-free direct lysis, isothermal amplification and prompt color development. The direct lysis process allows for release of genomic DNA within 2 min. The isothermal amplification reaction exhibits excellent analytical performance, detecting Pinelliae Rhizoma at levels as low as 0.1 pg and Rhizoma Typhonii Flagelliformis at 0.08 ng, while also demonstrating good linearity across a wide range, making it suitable for quantitative analysis. Two different color reagents are employed for these two different plant materials, resulting in dual-color changes for captivating visual detection. The developed direct-LAMP was also successfully validated across 48 actual samples.

摘要

半夏以其对咳嗽、失眠和炎症的药用功效而闻名,常用于植物性膳食补充剂和草药产品中。然而,由于需求旺盛,存在掺假风险。本研究引入了直接环介导等温扩增技术(Direct-LAMP),这是一种快速比色技术,用于区分半夏与其主要掺假品水半夏。该方法包括三个部分:无需DNA提取的直接裂解、等温扩增和快速显色。直接裂解过程可在2分钟内释放基因组DNA。等温扩增反应具有出色的分析性能,可检测低至0.1 pg的半夏和0.08 ng的水半夏,同时在很宽的范围内也表现出良好的线性,适用于定量分析。针对这两种不同的植物材料使用了两种不同的显色试剂,产生双色变化以实现引人注目的视觉检测。所开发的直接LAMP方法也在48个实际样品中成功得到验证。

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