Specific and Sensitive Detection of Escherichia coli O157:H7 in Meat by a Multiplex PCR

Abstract:

A multiplex PCR assay for detection of Escherichia coli O157:H7 was developed by using 3 sets of primers that specifically amplify segments of the rfbE、fliC and eaeA genes. The target genes fragment of the PCR assay were 291 bp, 625 bp and 368 bp, respectively. Analysis of 30 strains demonstrated that this PCR system was specific. The detection limit of the PCR was 91.35 pg with genomic DNA. This multiplex PCR assay did not cross-react with the background Salmonella?typhimurium and could detect 1.4 CFU/mL in artificial inoculated meat sample after enrichment at 37℃ for 6 h. Three of 30 meat samples were detected positive. These results indicated that the multiplex PCR assay can be used for specific and sensitive detection of E. coli O157:H7.

Key words:Escherichia coli O157:H7, Multiplex PCR, Meat