title: Enterohemorrhagic Escherichia coli as causes of hemolytic uremic syndrome in the Czech Republic.
abstract: 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.
  
  
  RESULTS
  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(H250]. 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. espPα encoding serine protease EspPα was found in EHEC O157:H7, O26:H11/NM, and O145:H28, whereas O172:NM and Orough:NM strains contained espPγ. 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
  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.
authors.affiliation: National Reference Laboratory for E. coli and Shigella, National Institute of Public Health, Prague, Czech Republic ; The 3rd Medical Faculty, Charles University Prague, Prague, Czech Republic.
authors.forename: Monika
authors.initials: M
authors.lastname: Marejková
authors.forename: Květa
authors.initials: K
authors.lastname: Bláhová
authors.forename: Jan
authors.initials: J
authors.lastname: Janda
authors.forename: Angelika
authors.initials: A
authors.lastname: Fruth
authors.forename: Petr
authors.initials: P
authors.lastname: Petráš
mesh_terms: D000900:Anti-Bacterial Agents / Q000494:pharmacology; D002675:Child, Preschool; D018153:Czech Republic / Q000453:epidemiology; D054324:Enterohemorrhagic Escherichia coli / Q000145:classification* / Q000187:drug effects / Q000235:genetics / Q000302:isolation & purification; D005260:Female; D005838:Genotype; D006463:Hemolytic-Uremic Syndrome / Q000175:diagnosis / Q000453:epidemiology* / Q000382:microbiology*; D006801:Humans; D007223:Infant; D008297:Male; D008826:Microbial Sensitivity Tests; D058885:Multilocus Sequence Typing; D010641:Phenotype; D010802:Phylogeny; D012621:Seasons; D012703:Serotyping; D022621:Shiga Toxin / Q000235:genetics; D014774:Virulence / Q000235:genetics
chemical_list: D000900:Anti-Bacterial Agents; D022621:Shiga Toxin
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