Establishment and application of a triple LAMP assay for rapid detection of three major foodborne pathogens in aquatic products
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Abstract
In the report, the specific genes of Vibrio parahaemolyticus, V. vulnificus, and Listeria monocytogenes were used as the targets, a triple visual Loop-mediated isothermal amplification (LAMP) assay was developed, the method performed the rapid detection at a constant temperature of 58 ℃ within 30 min, with a sensitivity of 100 fg·µL−1, and demonstrated the high specificity by distinguishing these pathogens from other common bacteria. The results can be visually interpreted based on color changes, and the pathogen identification was 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%-90.0%). In conclusion, a rapid, sensitive, and specific triple visual LAMP assay for detecting V. parahaemolyticus, V. vulnificus, and L. monocytogenes was successfully established, which provide an efficient screening tool for aquatic product safety monitoring.
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