Triplex PCR for Rapid Detection of Escherichia coli O157:H7 Directly from Ground Pork

A polymerase chain reaction (PCR) procedure is being developed to detect Escherichia coli O157:H7 and associated Shiga-like toxins (STX1 and STX2) directly from pork, bypassing the time-consuming microbial culture steps for identification. A triplex PCR procedure was adapted and conditions experimentally determined to efficiently amplify three target genes: uidA with unique sequence in E. coli O157:H7, and stx1 and stx2. A meat sample preparation technique developed in previous research was modified to allow detection of 0.2-2.0 colony forming units per gram (CFU./g.) of meat. Time required to complete the procedure after overnight incubation of samples is seven hours