Establishment of a multiplex PCR detection method for three foodborne pathogens

Objective:To establish a rapid multiplex polymerase chain reaction(PCR) method for the simultaneous detection of Salmonella spp.,Shigella spp. and Enterohemorrhagic Escherichia coli O157∶H7. Methods:Three pairs of primers had been designed based on the invA gene in Salmonella spp.,ipaH gene in Shigella spp.and uidA gene in Enterohemorrhagic Escherichia coli O157∶H7,single-factor experiment and L9(34) orthogonal experiment were used to optimize multiplex PCR amplification system,and the sensitivity of multiplex PCR was also analyzed. Results:Fragments of 495,620,252 bp were obtained respectively. Under the optimized condition,the detection sensitivity of multiplex PCR for three pathogens was 104CFU/mL. This method had been applied to artificial experiment,the result was accurate and steady,which could be obtained within 5h.Conclusion:A simple,specific and sensitive multiplex PCR method had been established and it was valuable for rapid monitoring and diagnosis for Salmonella spp.,Shigella spp. and Enterohemorrhagic Escherichia coli O157∶H7.