Polynucleotide primers and probes for the specific detection of E. coli 0157:H7 and/or E. coli 0157:NM which produces verotoxin in samples are provided. The primers and probes can be used in real time diagnostic assays for rapid detection of E. coli 0157:H7 and/or E. coli 0157:NM which produces verotoxin. The primers and probes can be used in real-time diagnostic assays for rapid detection of E. coli 0157:H7 and/or E. coli 0157:NM which produces verotoxin in a variety of situations and are capable of distinguishing E. coli 0157:H7 and/or E. coli 0157:NM which produces verotoxin from other E. coli strains. Kits comprising the primers and probes are also provided.
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