Typing of O26 enterohaemorrhagic and enteropathogenic Escherichia coli isolated from humans and cattle with IS621 multiplex PCR-based fingerprinting

J Appl Microbiol. 2011 Sep;111(3):773-86. doi: 10.1111/j.1365-2672.2011.05089.x. Epub 2011 Jul 20.

Abstract

Aims: This study evaluated a typing method of O26:H11 enterohaemorrhagic and enteropathogenic Escherichia coli (EHEC and EPEC) based on the variation in genomic location and copy numbers of IS621.

Methods and results: Two multiplex PCRs, targeting either the left (5') or right (3') IS/chromosome junction of 12 IS621 insertion sites and one PCR specific of another truncated copy, were developed. Thirty-eight amplification profiles were observed amongst a collection of 69 human and bovine O26:H11 EHEC and EPEC. Seventy-one per cent of the 45 EHEC and EPEC with identical IS621 fingerprints within groups of two, three or four isolates had >85% pulsed field gel electrophoresis (PFGE) profile similarity, including four groups of epidemiologically related EHEC or EPEC, while most of the groups had <85% similarity between each others. Epidemiologically related EHEC from each of three independent outbreaks in Japan and Belgium also exhibited identical IS621 fingerprints and PFGE profiles.

Conclusions: The IS621 fingerprinting and the PFGE are complementary typing assays of EHEC and EPEC; though, the former is less discriminatory.

Significance and impact of the study: The IS621 printing method represents a rapid (24 h) first-line surveillance and typing assay, to compare and trace back O26:H11 EHEC and EPEC during surveys in farms, multiple human cases and outbreaks.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Bacterial Typing Techniques / methods*
  • Belgium
  • Cattle
  • DNA Fingerprinting / methods*
  • DNA, Bacterial / genetics
  • Electrophoresis, Gel, Pulsed-Field
  • Enterohemorrhagic Escherichia coli / classification*
  • Enterohemorrhagic Escherichia coli / genetics
  • Enterohemorrhagic Escherichia coli / isolation & purification
  • Enteropathogenic Escherichia coli / classification*
  • Enteropathogenic Escherichia coli / genetics
  • Enteropathogenic Escherichia coli / isolation & purification
  • Humans
  • Japan
  • Multiplex Polymerase Chain Reaction / methods*
  • O Antigens / genetics
  • Sequence Analysis, DNA

Substances

  • DNA, Bacterial
  • O Antigens