Real-time Fluorescence and Visual Colorimetric Loop-Mediated Isothermal Amplification Assays for the Rapid and Visual Identification of the Genus Diodon

文献类型: 外文期刊

第一作者: Xie, Ruibin

作者: Xie, Ruibin;Gao, Jie;Li, Hui;Yu, Wenjie;Zhang, Juan;Wang, Nan;Chen, Ailiang

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关键词: Diodon; Loop-mediated isothermal amplification; Pufferfish species identification; Food safety; Visual detection; Fluorescence detection

期刊名称:FOOD ANALYTICAL METHODS ( 影响因子:3.498; 五年影响因子:3.226 )

ISSN: 1936-9751

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收录情况: SCI

摘要: In the present study, real-time fluorescence loop-mediated isothermal amplification (flo-LAMP) and visual colorimetric (vis-LAMP) assays were established for Diodon identification. LAMP primers were designed for the Diodon cytochrome oxidase I gene, and their high specificity was validated against 21 other fish species. LAMP has a minimum detection limit of 0.5 pg/mu L for genomic DNA. Furthermore, LAMP identified Diodon in both cooked and digested samples, with the results showing high application potential. The set of LAMP assays was carried out within 1 h under constant temperature conditions. For the flo-LAMP, a positive result was judged by an S-shaped amplification curve. For the vis-LAMP, the results were observed with the naked eye during daylight. Therefore, these rapid, reliable, sensitive, and inexpensive Diodon identification methods could be used to monitor the presence of fish from the Diodon genus in the food industry and detect cases of Diodon poisoning.

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