title: Characterization of fluoroquinolone and cephalosporin resistance mechanisms in Enterobacteriaceae isolated in a Dutch teaching hospital reveals the presence of an Escherichia coli ST131 clone with a specific mutation in parE.
abstract: OBJECTIVE
  To characterize the mechanisms of fluoroquinolone and cephalosporin resistance in Enterobacteriaceae from a Dutch teaching hospital in 2008.
  
  
  METHODS
  We sequenced gyrA, gyrB, parC and parE. The presence of plasmid-encoded genes qnrA, qnrB, qnrS, aac(6')-Ib, qepA, bla(TEM), bla(SHV,) bla(OXA), bla(CTX-M) and bla(AmpC) was studied by PCR. Escherichia coli isolates were further characterized by AFLP and multilocus sequence typing (MLST).
  
  
  RESULTS
  In total, 49 E. coli, 16 Klebsiella pneumoniae and 3 Enterobacter cloacae isolates were investigated. Mutations in gyrA were found in all E. coli isolates. Forty-five (92%) E. coli isolates carried at least one point mutation in parC. Most E. coli isolates (59%) also carried mutations in parE, of which I529L was the most prevalent. I529L was unequivocally associated with E. coli sequence type (ST) 131. This single-nucleotide polymorphism (SNP) was later also found in eight out of nine ST131 strains from another collection. Twenty-nine E. coli isolates carried extended-spectrum β-lactamase (ESBL) genes, predominantly bla(CTX-M-15). In E. coli, aac(6')-Ib-cr was the predominant plasmid-mediated resistance mechanism, whereas in K. pneumoniae qnr genes were found mostly. In K. pneumoniae isolates, qnr and aac(6')-Ib-cr co-occurred with ESBL genes (n = 13; bla(CTX-M) and bla(SHV)) and/or bla(AmpC) (n = 3; bla(DHA-1)).
  
  
  CONCLUSIONS
  E. coli ST131 was the predominant clone, which accumulated a high number of chromosomal mutations. The I529L SNP in parE was a signature of most, but not all, ST131 strains. In contrast to E. coli, fluoroquinolone resistance mechanisms were predominantly plasmid-encoded in K. pneumoniae.
authors.affiliation: Department of Clinical Microbiology, Leiden University Medical Center, Leiden, The Netherlands. s.paltansing@lumc.nl
authors.forename: S
authors.initials: S
authors.lastname: Paltansing
authors.forename: M E M
authors.initials: ME
authors.lastname: Kraakman
authors.forename: J M C
authors.initials: JM
authors.lastname: Ras
authors.forename: E
authors.initials: E
authors.lastname: Wessels
authors.forename: A T
authors.initials: AT
authors.lastname: Bernards
mesh_terms: D018550:Cephalosporin Resistance / Q000187:drug effects / Q000235:genetics*; D003001:Cloning, Molecular; D027101:DNA Topoisomerase IV / Q000737:chemistry / Q000235:genetics*; D004755:Enterobacteriaceae / Q000187:drug effects / Q000235:genetics / Q000302:isolation & purification; D004926:Escherichia coli / Q000187:drug effects / Q000235:genetics* / Q000302:isolation & purification; D024841:Fluoroquinolones / Q000494:pharmacology; D006784:Hospitals, Teaching; D006801:Humans; D008826:Microbial Sensitivity Tests / Q000379:methods; D009154:Mutation / Q000235:genetics*; D009426:Netherlands / Q000453:epidemiology; D020641:Polymorphism, Single Nucleotide / Q000235:genetics
chemical_list: D024841:Fluoroquinolones; D027101:DNA Topoisomerase IV