Establishment and application of a triple LAMP assay for rapid detection of three major foodborne pathogens in aquatic products

Vibrio parahaemolyticus, V. vulnificus, and Listeria monocytogenes are three common foodborne pathogens in aquatic products, posing significant threats to human health through single or mixed infections. In this study, a triple visual Loop-mediated isothermal amplification (LAMP) assay was developed, targeting specific genes of these pathogens. The method enables rapid detection at a constant temperature of 58°C within 30 min, with a sensitivity of 100 fg·µL⁻¹, and demonstrates high specificity by distinguishing these pathogens from other common bacteria. Results can be visually interpreted based on color changes, and pathogen identification is further confirmed by restriction enzyme digestion. When applied to 40 randomly collected shellfish samples (20 oysters and 20 mussels), the assay achieved a 100% detection rate, significantly outperforming triple PCR (72.5%-90%). In conclusion, this study successfully established a rapid, sensitive, and specific triple visual LAMP assay for detecting V. parahaemolyticus, V. vulnificus, and L. monocytogenes, providing an efficient screening tool for aquatic product safety monitoring.