title: Emergence of a multiresistant KPC-3 and VIM-1 carbapenemase-producing Escherichia coli strain in Spain.
abstract: OBJECTIVE
  To characterize the mechanisms involved in carbapenem resistance, as well as the genetic elements supporting their mobilization, in a multidrug-resistant Escherichia coli isolate.
  
  
  METHODS
  The E. coli isolate was obtained from a patient with fatal urinary sepsis. Antimicrobial susceptibility testing was performed by the disc diffusion and agar dilution methods. The E. coli molecular type and phylogroup were determined using multilocus sequence typing and the triple PCR technique, respectively. PCR and sequencing were used for virulence and resistance genotype characterization. Plasmid content and gene location were analysed by S1-PFGE, I-Ceu1-PFGE and hybridization experiments. Transformation assays were performed.
  
  
  RESULTS
  The E. coli strain, typed as ST448 and phylogroup B1, was resistant to all tested antibiotics except fosfomycin, tigecycline and tetracycline. The following resistance and virulence genetic structures were obtained: ISKpn7 + bla(KPC-3) + ISKpn6 linked to Tn4401; tnpR + aac(6')-Ib'-9 + aadA1 + bla(OXA-9) + tnpR + bla(TEM-1a) + tnpB + strB + strA + sul2; intI1 + bla(VIM-1) + aac(6')-Ib' + aphA15 + aadA1 + catB2 + qacEΔ1-sul1 + orf5; ISEcp1 + bla(CMY-2); IS26 + bla(SHV-12); aph(3')-I; aac(3)-IV; floR; catA; and fimA. Mutations in the ampC promoter (-18, -1 and +58) and substitutions in the GyrA (Ser-83→Leu and Asp-87→Asn) and ParC (Ser-80→Ile) proteins were observed. IncFII (ST2), IncA/C and ColE(TP) plasmids of 145.5, 87 and <2 kb, respectively, were found. The bla(VIM-1) gene was located in a non-typeable plasmid of >300 kb, and the bla(KPC-3) gene in the 145.5 kb IncFII plasmid. Transformant strains carried the IncFII and ColE(TP) plasmids, and the bla(KPC-3), bla(TEM-1a), bla(OXA-9), aadA1, aac(6')-Ib'-9, aac(3)-IV and floR genes.
  
  
  CONCLUSIONS
  This is the first report of the co-production of KPC-3, VIM-1, SHV-12, OXA-9 and CMY-2 in a unique clinical multiresistant E. coli isolate. The dissemination of these genes on mobile genetic elements is alarming and complicates antimicrobial therapies.
authors.affiliation: Área de Bioquímica y Biología Molecular, Universidad de La Rioja, Logroño, Spain Área de Microbiología Molecular, Centro de Investigación Biomédica de La Rioja (CIBIR), Logroño, Spain.
authors.forename: Nerea
authors.initials: N
authors.lastname: Porres-Osante
authors.affiliation: Departamento de Diagnóstico Biomédico, Laboratorio de Microbiología, Hospital San Pedro, Logroño, Spain.
authors.forename: Jose Manuel
authors.initials: JM
authors.lastname: Azcona-Gutiérrez
authors.affiliation: Área de Microbiología Molecular, Centro de Investigación Biomédica de La Rioja (CIBIR), Logroño, Spain.
authors.forename: Beatriz
authors.initials: B
authors.lastname: Rojo-Bezares
authors.affiliation: Departamento de Diagnóstico Biomédico, Laboratorio de Microbiología, Hospital San Pedro, Logroño, Spain.
authors.forename: Esther
authors.initials: E
authors.lastname: Undabeitia
authors.affiliation: Área de Bioquímica y Biología Molecular, Universidad de La Rioja, Logroño, Spain Área de Microbiología Molecular, Centro de Investigación Biomédica de La Rioja (CIBIR), Logroño, Spain.
authors.forename: Carmen
authors.initials: C
authors.lastname: Torres
authors.affiliation: Área de Microbiología Molecular, Centro de Investigación Biomédica de La Rioja (CIBIR), Logroño, Spain ysaenz@riojasalud.es.
authors.forename: Yolanda
authors.initials: Y
authors.lastname: Sáenz
mesh_terms: D000368:Aged; D001426:Bacterial Proteins / Q000235:genetics* / Q000378:metabolism*; D004269:DNA, Bacterial / Q000737:chemistry / Q000235:genetics; D024901:Drug Resistance, Multiple, Bacterial; D004926:Escherichia coli / Q000187:drug effects* / Q000201:enzymology* / Q000235:genetics / Q000302:isolation & purification; D004927:Escherichia coli Infections / Q000382:microbiology*; D017809:Fatal Outcome; D005260:Female; D022761:Gene Transfer, Horizontal; D006801:Humans; D020071:Interspersed Repetitive Sequences; D008826:Microbial Sensitivity Tests; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D017422:Sequence Analysis, DNA; D013030:Spain; D014552:Urinary Tract Infections / Q000382:microbiology; D001618:beta-Lactamases / Q000235:genetics* / Q000378:metabolism*
chemical_list: D001426:Bacterial Proteins; D004269:DNA, Bacterial; D001618:beta-Lactamases; C063912:carbapenemase
keywords: CMY-2; ST448; aac(6′)-Ib′; carbapenems; plasmids