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

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.

Automated summary

Real-time fluorescence loop-mediated isothermal amplification (flo-LAMP) and visual colorimetric (vis-LAMP) assays were established for Diodon identification. These methods are rapid, reliable, sensitive, and inexpensive, and can be used to monitor the presence of fish from the Diodon genus in the food industry and detect cases of Diodon poisoning.