title: Molecular epidemiology of extended-spectrum-β-lactamase-producing Klebsiella pneumoniae over a 10 year period in Calgary, Canada.
abstract: OBJECTIVE
  A study was designed to investigate the molecular epidemiology of extended-spectrum β-lactamase (ESBL)-producing Klebsiella pneumoniae isolated in a centralized region over a 10 year period (2000-09).
  
  
  METHODS
  Molecular characterization was done using isoelectric focusing, PCR and sequencing for bla(CTX-M), bla(TEM) and bla(SHV) genes and plasmid-mediated quinolone resistance determinants. Genetic relatedness was determined with PFGE using XbaI and multilocus sequencing typing.
  
  
  RESULTS
  A total of 89 patients with incident infections were identified; the majority presented with hospital-onset urinary tract infections. The absolute number of ESBL-producing isolates remained very low until 2003, increased slightly in 2004, remained stable until 2008 and then in 2009 there was an abrupt increase in the numbers of ESBL producers identified. The majority of K. pneumoniae produced CTX-M-14 and -15, and have replaced SHV-12-producing isolates since 2005. We identified four different major sequence types (STs) among 32% of isolates (i.e. ST17, ST20, and the new ST573 and ST575) and provided insight into their clinical and molecular characteristics. The ST isolates were more likely to produce community-onset infections, were associated with bla(CTX-M) and emerged during the latter part of the study period. ST17 produced CTX-M-15 and SHV-12, and was more likely to be positive for qnrB; ST20 produced CTX-M-14 and was positive for qnrS. The multiresistant ST575 that produced CTX-M-15 appeared in 2009.
  
  
  CONCLUSIONS
  Our study highlights the importance of molecular epidemiology in providing insight into the emergence, characteristics and distribution of STs among ESBL-producing K. pneumoniae.
authors.affiliation: Division of Microbiology,  Calgary Laboratory Services, Calgary, Alberta,  Canada.
authors.forename: Gisele
authors.initials: G
authors.lastname: Peirano
authors.forename: Jessica Hung King
authors.initials: JH
authors.lastname: Sang
authors.forename: André
authors.initials: A
authors.lastname: Pitondo-Silva
authors.forename: Kevin B
authors.initials: KB
authors.lastname: Laupland
authors.forename: Johann D D
authors.initials: JD
authors.lastname: Pitout
mesh_terms: D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D002170:Canada / Q000453:epidemiology; D016000:Cluster Analysis; D005260:Female; D005838:Genotype; D006801:Humans; D007525:Isoelectric Focusing; D007710:Klebsiella Infections / Q000453:epidemiology* / Q000382:microbiology*; D007711:Klebsiella pneumoniae / Q000145:classification / Q000201:enzymology* / Q000235:genetics / Q000302:isolation & purification*; D008297:Male; D008875:Middle Aged; D017720:Molecular Epidemiology; D058889:Molecular Typing; D016133:Polymerase Chain Reaction; D017422:Sequence Analysis, DNA; D001618:beta-Lactamases / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*
chemical_list: D001618:beta-Lactamases