Molecular characterisation of Escherichia coli O157:H7 isolates by pulsed-field gel electrophoresis and plasmid DNA analysis.

Abstract

Foods of bovine origin have been identified as sources of Escherichia coli O157:H7. Genomic DNA of E. coli O157:H7 isolates from patients (six isolates), food samples (18 isolates from ground beef and six isolates from raw milk) and calf faecal samples (31 isolates) were characterised by pulsed-field gel electrophoresis (PFGE) and plasmid DNA analysis. These isolates originated from different locations in the USA during 1992 and 1993. Twenty-one distinct genomic profiles were generated from the 61 isolates by PFGE after digestion with the endonuclease XbaI. Four genomic profiles were identified among five patient isolates and the remaining patient isolate was not typable. Five different profiles were detected amongst the isolates from ground beef, one of which was associated with 13 ground beef isolates from an outbreak in the Pacific Northwest of the USA in 1993. The PFGE profile of five calf isolates from Washington and Wisconsin was identical to the profile of the ground beef isolates from the outbreak, suggesting that these isolates were related. Similarly, one PFGE profile accounted for three isolates from calf faeces and one from ground beef. Six raw milk isolates from Georgia were indistinguishable both from each other and from one isolate from calf faeces. Fourteen genomic profiles were identified among 31 calf faecal isolates from 18 different herds in 11 states. Only five plasmid profiles were identified among the 61 isolates. PFGE was shown to be a useful typing technique for E. coli O157:H7.