Probe-based Loop-Mediated Isothermal Amplification Assay for Multi-target Quantitative Detection of Three Foodborne Pathogens in Seafood

Foodborne pathogens pose serious risks to human health and food safety. More than one pathogen exists in seafood. It is difficult for conventional culture methods to detect some foodborne pathogens simultaneously. Probe-based loop-mediated isothermal amplification is a feasible tool for achieving multi-objective detection of bacterial pathogens in a short time. Here we present a simultaneous detection method of Staphylococcus aureus, Vibrio parahaemolyticus, and Salmonella spp. for microbiological monitoring. With a sensitivity of low to 101 CFU/mL of bacteria, the assay can be used in the identification of three bacterial target fragments in a single reaction within 25 min. No cross-reactivity was observed. After enrichment for 18 h, the limit of MFP-LAMP assay detection in spiked foods was 5 CFU/25 g. In the light of the obtained results of the study, the probe-based LAMP assay provides an efficient and rapid approach to monitoring foodborne disease for its early diagnosis without being time-consuming and labor-intensive. It is important to both protect consumer health and obtain safe and quality seafood.