Establishment of a multiplex PCR method for rapid detection of two kinds of diarrheagenic Escherichia coli

To establish a rapid multiplex PCR method, which can simultaneously detect Enterohemorrhagic Escherichia coli O157∶ H7 and Enteroinvasive Escherichia coli, three pairs of primers had been designed based on the O- antigen gene in Enterohemorrhagic Escherichia coli O157 ∶ H7, ipa H gene in Enteroinvasive Escherichia coli and uid A gene in all Escherichia coli.The method was established and optimized, and the specificity and sensitivity were also analyzed. This method was applied to the detection of the artificially infected fresh lettuce with two pathogens.The results showed that the triplex PCR could detect two kinds of diarrheagenic Escherichia coli. The sensitivity was 6.3 × 103 CFU / m L.This method had high specificity and sensitivity.The sensitivity of the detection of artificial lettuce infected with two pathogens was 7.8 × 104 CFU / m L.It was suggested that the method was suitable for rapid detection of Enterohemorrhagic Escherichia coli O157 ∶ H7 and Enteroinvasive Escherichia coli in fresh agricultural products.