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Applied and Environmental Microbiology, January 2007, p. 22-31, Vol. 73, No. 1
0099-2240/07/$08.00+0     doi:10.1128/AEM.00982-06
Copyright © 2007, American Society for Microbiology. All Rights Reserved.

Identification of Escherichia coli O157:H7 Genomic Regions Conserved in Strains with a Genotype Associated with Human Infection{triangledown} ,{dagger}

Marina Steele,1 Kim Ziebell,1 Yongxiang Zhang,2 Andrew Benson,3 Paulina Konczy,1 Roger Johnson,1 and Victor Gannon2*

Laboratory for Foodborne Zoonoses, Public Health Agency of Canada, Guelph, Ontario, Canada N1G 3W4,1 Laboratory for Foodborne Zoonoses, Public Health Agency of Canada, Lethbridge, Alberta, Canada T1J 3Z4,2 Department of Food Science and Technology, University of Nebraska, Lincoln, Nebraska 68583-09193

Received 26 April 2006/ Accepted 9 October 2006

Beta-glucuronidase-negative, sorbitol-nonfermenting isolates of Shiga toxin-producing Escherichia coli O157 comprise part of a clone complex of related enterohemorrhagic E. coli isolates. High-resolution genotyping shows that the O157 populations have diverged into two different lineages that appear to have different ecologies. To identify genomic regions unique to the most common human-associated genotype, suppression subtractive hybridization was used to identify DNA sequences present in two clinical strains representing the human lineage I O157:H7 strains but absent from two bovine-derived lineage II strains. PCR assays were then used to test for the presence of these regions in 10 lineage I strains and 20 lineage II strains. Twelve conserved regions of genomic difference for lineage I (CRDI) were identified that were each present in at least seven of the lineage I strains but absent in most of the lineage II strains tested. The boundaries of the lineage I conserved regions were further delimited by PCR. Eleven of these CRDI were associated with E. coli Sakai S-loops 14, 16, 69, 72, 78, 82, 83, 91 to 93, 153, and 286, and the final CRDI was located on the pO157 virulence plasmid. Several potential virulence factors were identified within these regions, including a putative hemolysin-activating protein, an iron transport system, and several possible regulatory genes. Cluster analysis based on lineage I conserved regions showed that the presence/absence of these regions was congruent with the inferred phylogeny of the strains.

* Corresponding author. Mailing address: Laboratory for Foodborne Zoonoses, Public Health Agency of Canada, 1st floor, C.F.I.A. Building, Lethbridge, AB T1J 3Z4, Canada. Phone: (403) 382-5514. Fax: (403) 381-1202. E-mail: gannonv{at}inspection.gc.ca.

{triangledown} Published ahead of print on 20 October 2006.

{dagger} Supplemental material for this article may be found at http://aem.asm.org/.

Applied and Environmental Microbiology, January 2007, p. 22-31, Vol. 73, No. 1
0099-2240/07/$08.00+0     doi:10.1128/AEM.00982-06
Copyright © 2007, American Society for Microbiology. All Rights Reserved.



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