Establishment of Multiple Fluorescence PCR Detection Method for Two Foodborne Pathogenic Bacteria
This study aims to establish a TaqMan dual fluorescent quantitative polymerase chain reaction(PCR)detection method for the rapid simultaneous detection of Salmonella and Vibrio parahaemolyticus in food.Specific detection primers and probes targeting the conservative sequences of the invA gene of Salmonella and the tlh gene of Vibrio parahaemolyticus were designed,and their concentrations and annealing temperatures were optimized to establish a dual-fluorescent quantitative PCR detection system,and evaluating the sensitivity,specificity and stability of the method,The results showed that the established dual fluorescence quantitative PCR detection method could specifically amplify Salmonella and Vibrio parahaemolyticus,while other strains did not amplify.The minimum detection value of Salmonella detection can reach 1 copies/μL,and the sensitivity of Vibrio parahaemolyticus detection is 10 copies/μL.The intra-batch and inter-batch coefficients of variation were both less than 2％,and the repeatability and stability were good,the results were consistent with those of traditional methods,and the time is shortened.In summary,the dual fluorescent PCR detection method established can quickly and accurately detect Salmonella and Vibrio parahaemolyticus in food,which can provide technical support for the rapid detection of foodborne pathogens bacteria in food.
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