This study aimed to establish a rapid detection method for Escherichia coli, Salmonella typhimurium, and Staphylococcus aureus using multiplex nested polymerase chain reaction (PCR) combined with capillary electrophoresis.Targeting the uidA gene of E.coli (β-glucuronidase), the hilA gene of S.typhimurium (virulence gene), and the nuc gene of S.aureus (thermostable nuclease gene), three pairs of nested primers were designed for two rounds of amplification.The PCR products were analyzed by capillary electrophoresis, and the annealing temperatures for the two rounds of nested PCR were optimized to establish a multiplex nested PCR-capillary electrophoresis detection method for the three foodborne pathogens.The three pairs of nested primers showed high specificity, with optimal annealing temperatures of 54 ℃ and 56 ℃ for the two rounds.The detection sensitivity reached 101 copies for E.coli and S.aureus, and 103 copies for S.typhimurium.This method is rapid, accurate, and highly sensitive, making it suitable for detecting these three foodborne pathogens in food samples from which DNA can be extracted, thereby providing a new approach for foodborne pathogen detection.
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