title: Complete sequences of a novel blaNDM-1-harbouring plasmid from Providencia rettgeri and an FII-type plasmid from Klebsiella pneumoniae identified in Canada.
abstract: OBJECTIVE
  Emergence of plasmids harbouring bla(NDM-1) is a major public health concern due to their association with multidrug resistance and their potential mobility.
  
  
  METHODS
  PCR was used to detect bla(NDM-1) from clinical isolates of Providencia rettgeri (PR) and Klebsiella pneumoniae (KP). Antimicrobial susceptibilities were determined using Vitek 2. The complete DNA sequence of two bla(NDM-1) plasmids (pPrY2001 and pKp11-42) was obtained using a 454-Genome Sequencer FLX. Contig assembly and gap closures were confirmed by PCR-based sequencing. Comparative analysis was done using BLASTn and BLASTp algorithms.
  
  
  RESULTS
  Both clinical isolates were resistant to all β-lactams, carbapenems, aminoglycosides, ciprofloxacin and trimethoprim/sulfamethoxazole, and susceptible to tigecycline. Plasmid pPrY2001 (113 295 bp) was isolated from PR. It did not show significant homology to any known plasmid backbone and contained a truncated repA and novel repB. Two bla(NDM-1)-harbouring plasmids from Acinetobacter lwoffii (JQ001791 and JQ060896) shared 100% similarity to a 15 kb region that contained bla(NDM-1). pPrY2001 also contained a type II toxin/antitoxin system. pKp11-42 (146 695 bp) was isolated from KP. It contained multiple repA genes. The plasmid backbone had the highest homology to the IncFIIk plasmid type (51% coverage, 100% nucleotide identity). The bla(NDM-1) region was unique in that it was flanked upstream by IS3000 and downstream by a novel transposon designated Tn6229. pKp11-42 also contained a number of mutagenesis and plasmid stability proteins.
  
  
  CONCLUSIONS
  pPrY2001 differed from all known plasmids due to its novel backbone and repB. pKp11-42 was similar to IncFIIk plasmids and contained a number of genes that aid in plasmid persistence.
authors.affiliation: Public Health Agency of Canada, Winnipeg, MB, Canada.
authors.forename: L F
authors.initials: LF
authors.lastname: Mataseje
authors.forename: D A
authors.initials: DA
authors.lastname: Boyd
authors.forename: B
authors.initials: B
authors.lastname: Lefebvre
authors.forename: E
authors.initials: E
authors.lastname: Bryce
authors.forename: J
authors.initials: J
authors.lastname: Embree
authors.forename: D
authors.initials: D
authors.lastname: Gravel
authors.forename: K
authors.initials: K
authors.lastname: Katz
authors.forename: P
authors.initials: P
authors.lastname: Kibsey
authors.forename: M
authors.initials: M
authors.lastname: Kuhn
authors.forename: J
authors.initials: J
authors.lastname: Langley
authors.forename: R
authors.initials: R
authors.lastname: Mitchell
authors.forename: D
authors.initials: D
authors.lastname: Roscoe
authors.forename: A
authors.initials: A
authors.lastname: Simor
authors.forename: G
authors.initials: G
authors.lastname: Taylor
authors.forename: E
authors.initials: E
authors.lastname: Thomas
authors.forename: N
authors.initials: N
authors.lastname: Turgeon
authors.forename: M R
authors.initials: MR
authors.lastname: Mulvey
mesh_terms: D000368:Aged; D002170:Canada; D004269:DNA, Bacterial / Q000737:chemistry / Q000235:genetics*; D004756:Enterobacteriaceae Infections / Q000382:microbiology; D005260:Female; D006801:Humans; D007711:Klebsiella pneumoniae / Q000201:enzymology* / Q000235:genetics* / Q000302:isolation & purification; D008297:Male; D008875:Middle Aged; D008969:Molecular Sequence Data; D010957:Plasmids; D011532:Providencia / Q000201:enzymology* / Q000235:genetics* / Q000302:isolation & purification; D017422:Sequence Analysis, DNA; D001618:beta-Lactamases / Q000235:genetics*
chemical_list: D004269:DNA, Bacterial; D001618:beta-Lactamases; C552864:beta-lactamase NDM-1
keywords: Enterobacteriaceae; carbapenemases; replicon type