Characterization of Clinical Vancomycin-Resistant Enterococcus faecium Isolated in Eastern Hungary.

OBJECTIVES: The aim of our study was to characterize and elicit the genetic relatedness of emerging vancomycin-resistant enterococci (VRE) isolated between 2012 and 2015 at a teaching hospital in Debrecen, Hungary. RESULTS: Altogether 43 nonduplicate vancomycin-resistant Enterococcus faecium (VREfm) clinical isolates were obtained. Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry was used for species identification. Isolates showed 100% resistance to ampicillin and ciprofloxacin while 81.4% were resistant to gentamicin. PCR analysis revealed the presence of VanB in 40 and VanA in 3 isolates. Among ace, agg, and esp virulence genes only esp was found in seven cases. Modified microtiter-plate test showed 13 weak and 4 moderate biofilm producer isolates. Pulsed-field gel electrophoresis revealed nine pulsotypes. According to multilocus sequence typing all of the tested isolates belonged to clonal complex 17 (CC17). CONCLUSIONS: We report on the alarming emergence of multidrug-resistant VREfm belonging to CC17 at a tertiary hospital in Eastern Hungary. This is the first report of sequence types 412 and 364 from this region. Although outbreak did not occur the increasing prevalence of VREfm is of concern and dissemination must be prevented with proper infection control measures and regular VRE screening.

Prevalence of vanC vancomycin-resistant enterococci in the teaching hospitals of the University of Debrecen, Hungary.

Vancomycin-resistant enterococci (VRE) are common nosocomial pathogens; however, until now they have been rarely encountered in Hungary. In the present study, we investigated the prevalence of VRE in the teaching hospitals of the University of Debrecen. Of 7,271 Enterococcus-containing clinical samples collected between 2004 and 2009, we identified 16 VRE. Species-specific polymerase chain reaction was used to detect Enterococcus faecalis, Enterococcus faecium, Enterococcus casseliflavus, and Enterococcus gallinarum. Multiplex polymerase chain reaction was performed to identify the vancomycin resistance genes: vanA, vanB, vanC1/C2, vanD, vanE, and vanG. Restriction digestion with SalI and HindIII was introduced to differentiate the vanC1 and vanC2 genes from each other. Genetic relationships between the strains were investigated by pulsed-field gel electrophoresis. Overall, we identified the vanC1 resistance gene in 14 E. gallinarum and the vanC2 resistance gene in two E. casseliflavus strains. Except for two samples, the isolates had different pulsed-field gel electrophoresis types, suggesting sporadic emergence of the resistant bacteria. In addition, antibiotic resistance profile was determined by E-test. Three E. gallinarum strains proved to be resistant to gentamicin because of the presence of the aacA-aphD gene. Although the prevalence of VRE in Debrecen is rather low, the appearance of multiple resistances is of concern.