title: Molecular epidemiology of carbapenem resistant Enterobacteriaceae in Valle d'Aosta region, Italy, shows the emergence of KPC-2 producing Klebsiella pneumoniae clonal complex 101 (ST101 and ST1789).
abstract: BACKGROUND
  The spread of carbapenem resistant Enterobacteriaceae (CRE) is an emerging clinical problem, of great relevance in Europe and worldwide. The aim of this study was the molecular epidemiology of CRE isolates in Valle d'Aosta region, Italy, and the mechanism of carbapenem resistance.
  
  
  RESULTS
  Sixty consecutive CRE samples were isolated from 52 hospital inpatients and/or outpatients from November 2013 to August 2014. Genotyping of microbial isolates was done by pulsed-field gel electrophoresis (PFGE) and multi-locus sequence typing (MLST), carbapenemases were identified by PCR and sequencing. Carbapenem resistance gene transfer was performed by filter mating, plasmids from parental and transconjugant strains were assigned to incompatibility groups by PCR-based replicon typing. Molecular characterization of CRE isolates assigned 25 Klebsiella pneumoniae isolates to PFGE types A1-A5 and sequencing type (ST) 101, 17 K. pneumoniae isolates to PFGE type A and ST1789 (a single locus variant of ST101), 7 K. pneumoniae isolates to PFGE types B or C and ST512, 2 K. pneumoniae isolates to PFGE type D and ST405, and 5 Escherichia coli isolates to PFGE type a and ST131. All K. pneumoniae ST101 and ST1789 isolates were extended-spectrum beta-lactamase (ESBL) producers and carried bla CTX-M-1 group gene; 4 K. pneumoniae ST101 isolates were resistant to colistin. Molecular analysis of beta-lactamase genes identified bla KPC-2 and bla CTX-M-group 1 into conjugative plasmid/s assigned to IncFII incompatibility group in ST101 and ST1789 K. pneumoniae isolates, bla KPC-3 into conjugative plasmid/s assigned to IncF incompatibility group in ST512 and ST405 K. pneumoniae isolates, bla VIM-1 into conjugative plasmid/s assigned to IncN incompatibility group in ST131 E. coli isolates.
  
  
  CONCLUSIONS
  The spread of CRE in Valle d'Aosta region was caused by the selection of KPC-2 producing K. pneumoniae ST101 and ST1789 epidemic clones belonging to clonal complex 101, KPC-3 producing K. pneumoniae epidemic clones assigned to ST512 and ST405, and VIM-1 producing E.coli ST131 epidemic clone. Carbapenem resistance, along with bla KPC-2, bla KPC-3 and bla VIM-1 carbapenemase genes, was transferred by conjugative plasmids assigned to IncFII, IncF, and IncN incompatibility groups, respectively, in filter mating experiments. The emergence of colistin resistance was observed in KPC-2 producing K. pneumoniae ST101 isolates.
authors.affiliation: Department of Public Health, University of Naples 'Federico II', Naples, Italy. mariateresa.88@libero.it.
authors.forename: Mariateresa
authors.initials: M
authors.lastname: Del Franco
authors.affiliation: Department of Public Health, University of Naples 'Federico II', Naples, Italy. laura.paone@libero.it.
authors.forename: Laura
authors.initials: L
authors.lastname: Paone
authors.affiliation: Medical Direction, Aosta Regional Hospital, Aosta, Italy. rnovati@ausl.vda.it.
authors.forename: Roberto
authors.initials: R
authors.lastname: Novati
authors.affiliation: Microbiology, Aosta Regional Hospital, Aosta, Italy. cgiacomazzi@ausl.vda.it.
authors.forename: Claudio G
authors.initials: CG
authors.lastname: Giacomazzi
authors.affiliation: Department of Public Health, University of Naples 'Federico II', Naples, Italy. maria.bagattini@unina.it.
authors.forename: Maria
authors.initials: M
authors.lastname: Bagattini
authors.affiliation: Medical Direction, Aosta Regional Hospital, Aosta, Italy. cgalott@ausl.vda.it.
authors.forename: Chiara
authors.initials: C
authors.lastname: Galotto
authors.affiliation: Microbiology, Aosta Regional Hospital, Aosta, Italy. pmontanera@ausl.vda.it.
authors.forename: Pier Giorgio
authors.initials: PG
authors.lastname: Montanera
authors.affiliation: Department of Public Health, University of Naples 'Federico II', Naples, Italy. triassi@unina.it.
authors.forename: Maria
authors.initials: M
authors.lastname: Triassi
authors.affiliation: Department of Public Health, University of Naples 'Federico II', Naples, Italy. rafzarri@unina.it.|CEINGE Biotecnologie Avanzate, Naples, Italy. rafzarri@unina.it.
authors.forename: Raffaele
authors.initials: R
authors.lastname: Zarrilli
mesh_terms: D000890:Anti-Infective Agents / Q000494:pharmacology; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism; D015373:Bacterial Typing Techniques; D015780:Carbapenems / Q000494:pharmacology; D024881:Drug Resistance, Bacterial / Q000235:genetics; D004755:Enterobacteriaceae / Q000187:drug effects / Q000201:enzymology / Q000235:genetics / Q000502:physiology*; D004756:Enterobacteriaceae Infections / Q000453:epidemiology* / Q000382:microbiology*; D005838:Genotype; D006801:Humans; D007558:Italy; D007711:Klebsiella pneumoniae / Q000187:drug effects / Q000201:enzymology / Q000235:genetics / Q000502:physiology*; D017720:Molecular Epidemiology; D010957:Plasmids / Q000235:genetics; D001618:beta-Lactamases / Q000235:genetics / Q000378:metabolism
chemical_list: D000890:Anti-Infective Agents; D001426:Bacterial Proteins; D015780:Carbapenems; D001618:beta-Lactamases; C063912:carbapenemase