title: Characterization of KPC-encoding plasmids from two endemic settings, Greece and Italy.
abstract: 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.
  
  
  
  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.
  
  
  
  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.
  
  
  
  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.
authors.affiliation: Department of Microbiology, National School of Public Health, Athens, Greece Department of Microbiology, Faculty of Medicine and University Hospital in Plzen, Charles University in Prague, Plzen, Czech Republic.
authors.forename: Costas C
authors.initials: CC
authors.lastname: Papagiannitsis
authors.affiliation: Department of Medical Biotechnologies, University of Siena, Siena, Italy Department of Surgery and Translational Medicine, University of Florence, Florence, Italy.
authors.forename: Vincenzo
authors.initials: V
authors.lastname: Di Pilato
authors.affiliation: Department of Medical Biotechnologies, University of Siena, Siena, Italy.
authors.forename: Tommaso
authors.initials: T
authors.lastname: Giani
authors.affiliation: Department of Microbiology, National School of Public Health, Athens, Greece.
authors.forename: Panagiota
authors.initials: P
authors.lastname: Giakkoupi
authors.affiliation: Department of Medical Biotechnologies, University of Siena, Siena, Italy Department of Experimental and Clinical Medicine, University of Florence, Florence, Italy.
authors.forename: Eleonora
authors.initials: E
authors.lastname: Riccobono
authors.affiliation: Department of Medical Biotechnologies, University of Siena, Siena, Italy.
authors.forename: Giulia
authors.initials: G
authors.lastname: Landini
authors.affiliation: Laboratory of Bacteriology, Hellenic Pasteur Institute, Athens, Greece.
authors.forename: Vivi
authors.initials: V
authors.lastname: Miriagou
authors.affiliation: Department of Microbiology, National School of Public Health, Athens, Greece Central Public Health Laboratory, Hellenic Centre of Disease Control and Prevention, Vari, Greece.
authors.forename: Alkiviadis C
authors.initials: AC
authors.lastname: Vatopoulos
authors.affiliation: Department of Medical Biotechnologies, University of Siena, Siena, Italy Department of Experimental and Clinical Medicine, University of Florence, Florence, Italy Microbiology and Virology Unit, Careggi University Hospital, Florence, Italy Don Carlo Gnocchi Foundation, Florence, Italy gianmaria.rossolini@unifi.it.
authors.forename: Gian Maria
authors.initials: GM
authors.lastname: Rossolini
mesh_terms: D001426:Bacterial Proteins / Q000235:genetics*; D015780:Carbapenems / Q000494:pharmacology; D024881:Drug Resistance, Bacterial / Q000235:genetics*; D016521:Electrophoresis, Gel, Pulsed-Field; D019353:Endemic Diseases; D058872:Epidemics; D004926:Escherichia coli / Q000235:genetics; D006115:Greece / Q000453:epidemiology; D006801:Humans; D007558:Italy / Q000453:epidemiology; D007710:Klebsiella Infections / Q000453:epidemiology / Q000382:microbiology*; D007711:Klebsiella pneumoniae / Q000187:drug effects / Q000201:enzymology / Q000235:genetics*; D008826:Microbial Sensitivity Tests; D058885:Multilocus Sequence Typing / Q000379:methods; D010957:Plasmids / Q000235:genetics*; D012150:Polymorphism, Restriction Fragment Length; D017422:Sequence Analysis, DNA; D001618:beta-Lactamases / Q000235:genetics*
chemical_list: D001426:Bacterial Proteins; D015780:Carbapenems; D001618:beta-Lactamases; C063912:carbapenemase