title: Plasmid comparison and molecular analysis of Klebsiella pneumoniae harbouring bla(KPC) from New York City and Toronto.
abstract: OBJECTIVE
  This study examined Klebsiella pneumoniae clinical isolates and their bla(KPC) plasmids to determine potential relatedness of the isolates and their plasmids harbouring carbapenem resistance mechanisms.
  
  
  METHODS
  K. pneumoniae carbapenemase (KPC)-producing K. pneumoniae from New York City (NYC) (n = 19) and Toronto (n = 2) were typed by PFGE and multilocus sequence typing (MLST). bla(KPC)-harbouring plasmids were transformed into Escherichia coli DH10B(TM), restricted using EcoRI and analysed for bla content and replicon (rep) type. Susceptibility profiles for clinical and transformed strains were determined by automated microbroth dilution using CLSI breakpoints. Outer membrane protein (OMP) genes were analysed by sequencing of ompk35 and ompk36.
  
  
  RESULTS
  PFGE analysis identified 17 related strains (≥ 80% similarity; 11 KPC-2, 6 KPC-3) where ST258 was the dominant clonal type. All clinical isolates contained both bla(SHV) and bla(TEM-1) and, with the exception of one isolate, were multidrug resistant (MDR). Transformed KPC plasmids (n = 21) carried TEM-1 (n = 18) and were MDR (n = 5). Three plasmid clusters, repFIIA (n = 10), repR (n = 3) and an unknown type (n = 3), were observed. repFllA plasmids were observed from both NYC and Toronto strains. OMP gene analysis revealed premature stop codons in ompk35 and numerous deletions and insertions in ompk36.
  
  
  CONCLUSIONS
  The dissemination of bla(KPC) is due both to carriage of similar KPC-harbouring plasmids within genetically distinct K. pneumoniae and to clonal spread of K. pneumoniae with unrelated KPC plasmids.
authors.affiliation: National Microbiology Laboratory, Winnipeg, Canada.
authors.forename: L F
authors.initials: LF
authors.lastname: Mataseje
authors.forename: D A
authors.initials: DA
authors.lastname: Boyd
authors.forename: B M
authors.initials: BM
authors.lastname: Willey
authors.forename: N
authors.initials: N
authors.lastname: Prayitno
authors.forename: N
authors.initials: N
authors.lastname: Kreiswirth
authors.forename: A
authors.initials: A
authors.lastname: Gelosia
authors.forename: S M
authors.initials: SM
authors.lastname: Poutanen
authors.forename: D E
authors.initials: DE
authors.lastname: Low
authors.forename: S G
authors.initials: SG
authors.lastname: Jenkins
authors.forename: K
authors.initials: K
authors.lastname: Katz
authors.forename: M R
authors.initials: MR
authors.lastname: Mulvey
mesh_terms: D000368:Aged; D000369:Aged, 80 and over; D000900:Anti-Bacterial Agents / Q000494:pharmacology; D001425:Bacterial Outer Membrane Proteins / Q000235:genetics; D015373:Bacterial Typing Techniques; D002170:Canada; D015780:Carbapenems / Q000494:pharmacology; D016521:Electrophoresis, Gel, Pulsed-Field; D005260:Female; D022761:Gene Transfer, Horizontal; D006801:Humans; D007710:Klebsiella Infections / Q000382:microbiology*; D007711:Klebsiella pneumoniae / Q000145:classification / Q000201:enzymology* / Q000235:genetics* / Q000302:isolation & purification; D008297:Male; D008826:Microbial Sensitivity Tests; D008875:Middle Aged; D017720:Molecular Epidemiology; D058889:Molecular Typing; D058885:Multilocus Sequence Typing; D009519:New York City; D010957:Plasmids; D014169:Transformation, Bacterial; D018440:beta-Lactam Resistance; D001618:beta-Lactamases / Q000235:genetics*
chemical_list: D000900:Anti-Bacterial Agents; D001425:Bacterial Outer Membrane Proteins; D015780:Carbapenems; D001618:beta-Lactamases