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Background  Enterohemorrhagic  Escherichia coli  (EHEC) cause diarrhea-associated hemolytic uremic syndrome (D+ HUS) worldwide, but no systematic study of EHEC as the causative agents of HUS was performed in the Czech Republic. We analyzed stools of all patients with D+ HUS in the Czech Republic between 1998 and 2012 for evidence of EHEC infection. We determined virulence profiles, phenotypes, antimicrobial susceptibilities and phylogeny of the EHEC isolates.    Methodology/Principal Findings  Virulence loci were identified using PCR, phenotypes and antimicrobial susceptibilities were determined using standard procedures, and phylogeny was assessed using multilocus sequence typing. During the 15-year period, EHEC were isolated from stools of 39 (69.4%) of 56 patients. The strains belonged to serotypes [ fliC  types] O157:H7/NM[ fliC   H7 ] (50% of which were sorbitol-fermenting; SF), O26:H11/NM[ fliC   H11 ], O55:NM[ fliC   H7 ], O111:NM[ fliC   H8 ], O145:H28[ fliC   H28 ], O172:NM[ fliC   H25 ], and Orough:NM[ fliC   H25 ]. O26:H11/NM[ fliC   H11 ] was the most common serotype associated with HUS (41% isolates). Five  stx  genotypes were identified, the most frequent being  stx   2a  (71.1% isolates). Most strains contained EHEC- hlyA  encoding EHEC hemolysin, and a subset (all SF O157:NM and one O157:H7) harbored  cdt-V  encoding cytolethal distending toxin.  espP α encoding serine protease EspPα was found in EHEC O157:H7, O26:H11/NM, and O145:H28, whereas O172:NM and Orough:NM strains contained  espP γ. All isolates contained  eae  encoding adhesin intimin, which belonged to subtypes β (O26), γ (O55, O145, O157), γ2/θ (O111), and ε (O172, Orough). Loci encoding other adhesins ( efa1, lpfA   O26 ,  lpfA   O157OI-141 ,  lpfA   O157OI-154 ,  iha ) were usually associated with particular serotypes. Phylogenetic analysis demonstrated nine sequence types (STs) which correlated with serotypes. Of these, two STs (ST660 and ST1595) were not found in HUS-associated EHEC before.    Conclusions/Significance  EHEC strains, including O157:H7 and non-O157:H7, are frequent causes of D+ HUS in the Czech Republic. Identification of unusual EHEC serotypes/STs causing HUS calls for establishment of an European collection of HUS-associated EHEC, enabling to study properties and evolution of these important pathogens.

Enterohemorrhagic Escherichia coli as Causes of Hemolytic Uremic Syndrome in the Czech Republic