title: Characterization of a highly glycopeptide-resistant Enterococcus gallinarum isolate.
abstract: OBJECTIVE
  Adequate treatment of emergency infection involving antibiotic-resistant bacteria such as vancomycin-resistant Enterococcus requires a convergence of clinical and bacteriologic techniques. An isolate of Enterococcus gallinarum, designated as TSGH63, is known to be uncommonly vancomycin-resistant. This study investigated the genetic determinant for this unique characteristic.
  
  
  METHODS
  After completing the conventional identification and sensitivity tests, the genomic content of E. gallinarum TSGH63 was extracted and analyzed by pulse-field electrophoresis. A set of specific primers for vanA, vanB, vanC1, and vanC2/C3 genes was then applied in a multiplex polymerase chain reaction (PCR) to differentiate its genetic content. To locate the determinant for high vancomycin resistance, the electrophoresis profile was further analyzed by Southern blot using the digoxigenin (DIG)-labeled vanA gene probe. Finally, interspecies transfer of the vancomycin-resistance determinant of E. gallinarum TSGH63 was tested by a conjugation experiment in vitro.
  
  
  RESULTS
  A 50-kb plasmid was identified in the analysis of the genomic extract of E. gallinarum TSGH63 by pulse field electrophoresis. Using multiplex PCR, we demonstrated that E. gallinarum TSGH63 harbors a vanA gene in addition to a vanC1 gene. The DIG-labeled vanA gene-specific probe bound to the plasmid exclusively on the Southern blot. The plasmid-carried vanA gene, but not the vanC1 gene, was found to be transferable from TSGH63 to E. faecalis JH2-2 by conjugation in vitro.
  
  
  CONCLUSIONS
  This is the first report of isolation of E. gallinarum with a high level of resistance to glycopeptides in Taiwan. The demonstrated interspecies transfer of the vancomycin-resistance gene highlights the importance of stringent control of the use of vancomycin.
authors.affiliation: Department of Pathology, Tri-Service General Hospital, Taipei, Taiwan.
authors.forename: J J
authors.initials: JJ
authors.lastname: Lu
authors.forename: J C
authors.initials: JC
authors.lastname: Wu
authors.forename: T S
authors.initials: TS
authors.lastname: Chiueh
authors.forename: C L
authors.initials: CL
authors.lastname: Perng
authors.forename: W M
authors.initials: WM
authors.lastname: Chi
authors.forename: W H
authors.initials: WH
authors.lastname: Lee
mesh_terms: D003227:Conjugation, Genetic; D016983:Enterococcus / Q000187:drug effects* / Q000235:genetics; D006801:Humans; D016133:Polymerase Chain Reaction; D020713:Vancomycin Resistance