title: Molecular epidemiology of high-level aminoglycoside-resistant enterococci isolated from patients in a university hospital in southern Italy.
abstract: OBJECTIVE
  We evaluated the genetic and molecular basis of high-level resistance to gentamicin and amikacin in 91 clinical isolates of Enterococcus faecalis and Enterococcus faecium in a university hospital in southern Italy from 1987 to 2003.
  
  
  METHODS
  Antibiotic susceptibility was evaluated by disc diffusion and microdilution methods. Genotyping was performed by PFGE and dendrogram analysis. Aminoglycoside resistance genes were analysed by multiplex PCR. Aminoglycoside resistance gene transfer was performed by filter mating.
  
  
  RESULTS
  In our strain collection, 44% of E. faecalis and 52% of E. faecium were high-level-resistant to gentamicin. Fifty-two PFGE profiles were identified for E. faecalis and 15 for E. faecium. Although the majority of PFGE patterns were single isolates, four patterns (two for E. faecalis and two for E. faecium) were isolated each in 8 and 4, and 6 and 4 different patients, respectively. The aac(6')-Ie-aph(2'')-Ia gene was responsible for high-level resistance to gentamicin and amikacin in E. faecalis and E. faecium; the aph(2'')-Id gene responsible for resistance to gentamicin was also isolated in E. faecium; the aph(3')-IIIa and ant(4')-Ia genes responsible for resistance to amikacin were also isolated in E. faecalis and E. faecium. High-level resistance to gentamicin, along with the aac(6')-Ie-aph(2'')-Ia gene, was transferred at a frequency of about 10(-5) to 10(-8) per recipient cell in 14 of 17 E. faecalis and 3 of 4 E. faecium different genotypes.
  
  
  CONCLUSIONS
  The spread of the aac(6')-Ie-aph(2'')-Ia gene was responsible for high-level resistance to gentamicin and amikacin among enterococci isolated from patients in our geographical area.
authors.affiliation: Dipartimento di Scienze Mediche Preventive, Università di Napoli 'Federico II', Naples, Italy. rafzarri@unina.it
authors.forename: Raffaele
authors.initials: R
authors.lastname: Zarrilli
authors.forename: Marie-Francoise
authors.initials: MF
authors.lastname: Tripodi
authors.forename: Anna
authors.initials: A
authors.lastname: Di Popolo
authors.forename: Rosaria
authors.initials: R
authors.lastname: Fortunato
authors.forename: Maria
authors.initials: M
authors.lastname: Bagattini
authors.forename: Margherita
authors.initials: M
authors.lastname: Crispino
authors.forename: Anna
authors.initials: A
authors.lastname: Florio
authors.forename: Maria
authors.initials: M
authors.lastname: Triassi
authors.forename: Riccardo
authors.initials: R
authors.lastname: Utili
mesh_terms: D000123:Acetyltransferases / Q000235:genetics; D000583:Amikacin / Q000494:pharmacology; D000617:Aminoglycosides / Q000494:pharmacology*; D000900:Anti-Bacterial Agents / Q000494:pharmacology; D002876:Chromosomes, Bacterial / Q000235:genetics / Q000378:metabolism; D004269:DNA, Bacterial / Q000235:genetics / Q000378:metabolism; D024881:Drug Resistance, Bacterial / Q000235:genetics*; D016521:Electrophoresis, Gel, Pulsed-Field; D013293:Enterococcus faecalis / Q000145:classification / Q000187:drug effects* / Q000235:genetics / Q000302:isolation & purification; D016984:Enterococcus faecium / Q000145:classification / Q000187:drug effects* / Q000235:genetics / Q000302:isolation & purification; D018014:Gene Transfer Techniques; D022761:Gene Transfer, Horizontal; D005798:Genes, Bacterial; D005838:Genotype; D005839:Gentamicins / Q000494:pharmacology; D016908:Gram-Positive Bacterial Infections / Q000382:microbiology*; D006785:Hospitals, University; D006801:Humans; D007558:Italy; D008826:Microbial Sensitivity Tests; D017720:Molecular Epidemiology; D017853:Phosphotransferases (Alcohol Group Acceptor) / Q000235:genetics; D016133:Polymerase Chain Reaction; D012150:Polymorphism, Restriction Fragment Length
chemical_list: D000617:Aminoglycosides; D000900:Anti-Bacterial Agents; D004269:DNA, Bacterial; D005839:Gentamicins; D000583:Amikacin; C108453:6'-aminoglycoside acetyltransferase-2''-aminoglycoside phosphotransferase; D000123:Acetyltransferases; D017853:Phosphotransferases (Alcohol Group Acceptor)