Characterization of KPC-encoding plasmids from two endemic settings, Greece and Italy

J Antimicrob Chemother. 2016 Oct;71(10):2824-30. doi: 10.1093/jac/dkw227. Epub 2016 Jun 21.

Abstract

Objectives: Global dissemination of KPC-type carbapenemases is mainly associated with the spread of high-risk clones of Klebsiella pneumoniae and of KPC-encoding plasmids. In this study, we explored the population structure of KPC-encoding plasmids from the recent epidemics of KPC-producing K. pneumoniae (KPC-Kp) in Greece and Italy, the two major European endemic settings.

Methods: Thirty-four non-replicate clinical strains of KPC-Kp representative of the early phases (2008-11) of the Greek (n = 22) and Italian (n = 12) epidemics were studied. Isolates were typed by MLST, and blaKPC-carrying plasmids were characterized by S1 profiling, PCR-based replicon typing and RFLP. Transfer experiments by conjugation or transformation were carried out with Escherichia coli recipients. Eleven plasmids, representative of all different restriction profiles, were completely sequenced.

Results: The representative Greek strains belonged to 14 sequence types (STs), with a predominance of ST258. The representative Italian strains belonged to three STs, with a predominance of clonal complex 258 (ST258, ST512). The 34 strains carried plasmids of variable size (78-166 kb), either with blaKPC-2 or blaKPC-3 gene embedded in a Tn4401a transposon. Plasmids from Greek strains were mostly of a single RFLP type (A) and resembled the archetypal pKpQIL KPC-encoding plasmid, while plasmids from Italian strains belonged to a more heterogeneous population, showing five RFLP profiles (A, C-F). Types A and C resembled pKpQIL or deletion derivatives thereof, while types D-F included plasmids with hybrid structures between pKpQIL, pKPN3 and pKPN101-IT.

Conclusions: pKpQIL-like plasmids played a major role in the dissemination of blaKPC in Greece and Italy, but evolved with different dynamics in these endemic settings.

MeSH terms

  • Bacterial Proteins / genetics*
  • Carbapenems / pharmacology
  • Drug Resistance, Bacterial / genetics*
  • Electrophoresis, Gel, Pulsed-Field
  • Endemic Diseases
  • Epidemics
  • Escherichia coli / genetics
  • Greece / epidemiology
  • Humans
  • Italy / epidemiology
  • Klebsiella Infections / epidemiology
  • Klebsiella Infections / microbiology*
  • Klebsiella pneumoniae / drug effects
  • Klebsiella pneumoniae / enzymology
  • Klebsiella pneumoniae / genetics*
  • Microbial Sensitivity Tests
  • Multilocus Sequence Typing / methods
  • Plasmids / genetics*
  • Polymorphism, Restriction Fragment Length
  • Sequence Analysis, DNA
  • beta-Lactamases / genetics*

Substances

  • Bacterial Proteins
  • Carbapenems
  • beta-Lactamases
  • carbapenemase