Loop-Mediated Isothermal Amplification for On-Site Visual Identification of Leech Species

IF 2.6 3区农林科学 Q2 FOOD SCIENCE & TECHNOLOGY Food Analytical Methods Pub Date : 2024-02-17 DOI:10.1007/s12161-024-02597-7

Jiangsong Peng, Ye Li, Xiaoli Deng, Mengyao Lu, Chunbin Yang, Yuping Shen, Guohua Xia, Huan Yang

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Abstract

Leeches are a well-known animal-derived health supplement commonly used as an anticoagulation and antithrombosis agent; however, adulteration and counterfeiting are often made for illegal profits. To identify leech species, this study developed a rapid, simple, and visualized method based on loop-mediated isothermal amplification (LAMP), which relies on a specific primer set designed according to the mitochondrial DNA control region of the target species. Quantitative polymerase chain reaction (qPCR) was also employed in parallel to compare the sensitivity and confirm the primer specificity. Primer sets with high specificity were successfully screened for LAMP reactions against four common leech species on the market. All of them have produced typical amplification profiles of the target sequences in qPCR reactions with significantly lower amplification sensitivity than LAMP assay. The newly established LAMP method in this study can be accomplished within 1 h, and it could be successfully applied for on-site visual identification of mislabeling and adulteration in the leech market.

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用于现场目测鉴定水蛭物种的环路介导等温扩增技术

水蛭是一种众所周知的动物源保健品，常用作抗凝血和抗血栓药物；然而，掺假和造假往往是为了非法牟利。为了鉴别水蛭的种类，本研究开发了一种基于环介导等温扩增（LAMP）的快速、简单和可视化方法，该方法依赖于根据目标物种线粒体 DNA 控制区设计的特定引物集。同时还采用了定量聚合酶链反应（qPCR）来比较灵敏度并确认引物的特异性。针对市场上四种常见水蛭物种的 LAMP 反应，成功筛选出了特异性较高的引物组。所有这些引物在 qPCR 反应中都能产生典型的目标序列扩增曲线，但扩增灵敏度明显低于 LAMP 方法。本研究新建立的 LAMP 方法可在 1 小时内完成，并可成功应用于水蛭市场中的现场目视识别错标和掺假。

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来源期刊

Food Analytical Methods 农林科学-食品科技

CiteScore

6.00

自引率

3.40%

发文量

244

审稿时长

3.1 months

期刊介绍： Food Analytical Methods publishes original articles, review articles, and notes on novel and/or state-of-the-art analytical methods or issues to be solved, as well as significant improvements or interesting applications to existing methods. These include analytical technology and methodology for food microbial contaminants, food chemistry and toxicology, food quality, food authenticity and food traceability. The journal covers fundamental and specific aspects of the development, optimization, and practical implementation in routine laboratories, and validation of food analytical methods for the monitoring of food safety and quality.