From June 2003 to December 2009, 98 isolates of vancomycin-resistant enterococci (VRE) were cultured from clinical specimens taken from patients admitted to a 1,500-bed tertiary-care teaching hospital in Beijing, China. Isolates were characterized by pulsed-field gel electrophoresis and multilocus sequence typing. We investigated the structure of the vanA gene cluster and the distribution of the virulence markers esp, hyl, gelE, asa1, and cylA by polymerase chain reaction. Our results indicate that multilocus sequence typing revealed five novel sequence types and one new allele. VRE faecium (VREfm) isolates were heterogeneous in their vanA cluster types and in the presence of virulence genes. We also observed inconsistency between genotype and phenotype in VREfm isolates. The outbreak with VREfm in our hospital appears polyclonal, whereas VRE faecalis characterization indicated dissemination of a particular clone. After 2007, VRE faecalis was completely replaced by VREfm, which has since been the predominant species in our hospital. VRE appears to be in an evolutionary flux in our hospital.
Get full access to this article
View all access options for this article.
References
1.
BrilliantovaA.N., KliasovaG.A., MironovaA.V., TishkovV.I., NovichkovaG.A., BobryninaV.O., SidorenkoS.V.2010. Spread of vancomycin-resistant Enterococcus faecium in two haematological centres in Russia. Int. J. Antimicrob. Agents, 35:177–181.
2.
CamargoI.L.B.C., ZanellaR.C., BrandileoneM.C.C., PignatariA.C.C., GoldmanG.H., WoodfordN., DariniA.L.C.2005. Occurrence of insertion sequences within the genomes and Tn1546-like elements of glycopeptide-resistant enterococci isolated in Brazil, and identification of a novel element, ISEfa5. Int. J. Med. Microbiol., 294:513–519.
3.
CaoB., LiuY.M., SongS.F., LiR., WangH., WangC.2008. First report of clinical and epidemiological characterization of vancomycin-resistant enterococci from mainland China. Int. J. Antimicrob. Agents, 32:279–281.
4.
Clinical, Laboratory Standards Institute. 2010. Performance standards for antimicrobial susceptibility testing. Twentieth informational supplement. Document M100–S20: CLSI, Wayne, PA.
5.
DepardieuF., PerichonB., CourvalinP.2004. Detection of the van alphabet and identification of Enterococci and Staphylococci at the species level by multiplex PCR. J. Clin. Microbiol., 42:5857–5860.
DuckW.M., StewardC.D., BanerjeeS.N., McGowanJ.E.Jr., TenoverF.C.2003. Optimization of computer software settings improves accuracy of pulsed-field gel electrophoresis macrorestriction fragment pattern analysis. J. Clin. Microbiol., 41:3035–3042.
8.
DupreI., ZanettiS., SchitoA.M., FaddaG., SechiL.A.2003. Incidence of virulence determinants in clinical Enterococcus faecium and Enterococcus faecalis isolates collected in Sardinia (Italy)J. Med. Microbiol., 52:491–498.
9.
GuL., CaoB., LiuY.M., GuoP., SongS.F., LiR.2009. A new Tn1546 type of VanB phenotype–vanA genotype vancomycin-resistant enterococcus faecium isolates in mainland China. Diagn. Microbiol. Infect. Dis., 63:70–75.
10.
HandwergerS., SkobleJ., DiscottoL.F., PucciM.J.1995. Heterogeneity of the vanA gene cluster in clinical isolates of enterococci from the northeastern United States. Antimicrob. Agents Chemother., 39:362–368.
11.
HashimotoY., TanimotoK., OzawaY., MurataT., IkeY.2000. Amino acid substitutions in the VanS sensor of the VanA-type vancomycin-resistant Enterococcus strains result in high-level vancomycin resistance and low level teicoplanin resistance. FEMS. Microbiol. Lett., 185:247–254.
12.
HidronA.I., wardsJ.R., PatelJ., HoranT.C., SievertD.M., PollockD.A., FridkinS.K.2008. NHSN annualupdate: antimicrobial-resistant pathogens associated with health care-associated infections: annual summary of data reported to the National Healthcare Safety Network at the Centers for Disease Control and Prevention, 2006–2007. Infect. Control Hosp. Epidemiol., 29:996–1011.
13.
HomanW.L., TribeD., PoznanskiS., LiM., HoggG., SpalburgE., van EmbdenJ.D.A., WillemsR.J. L.2002. Multilocus sequence typing scheme for Enterococcus faecium. J. Clin. Microbiol., 40:1963–1971.
14.
HsiehY.C., LeeW.S., OuT.Y., HsuehP.R.2010. Clonal spread of CC17 vancomycin-resistant Enterococcus faecium with multilocus sequence type 78 (ST78) and a novel ST444 in Taiwan. Eur. J. Clin. Microbiol. Infect. Dis., 29:25–30.
15.
KawalecM., KedzierskaJ., GajdaA., SadowyE., WegrzynJ., NaserS., SkotnickiA.B., GniadkowskiM., HryniewiczW.2007. Hospital outbreak of vancomycin-resistant enterococci caused by a single clone of Enterococcus raffinosus and several clones of Enterococcus faecium. Clin. Microbiol. Infect., 13:893–901.
16.
KlareI., KonstabelC., Mueller-BertlingS., WernerG., StrommengerB., KettlitzC., BorgmannS., SchulteB., JonasD., SerrA., FahrA.M., EignerU., WitteW.2005. Spread of ampicillin/vancomycin resistant Enterococcus faecium of the epidemic-virulent clonal complex-17 carrying the genes esp and hyl in German hospitals. Eur. J. Clin. Microbiol. Infect. Dis., 24:815–825.
17.
LauderdaleT.L., McDonaldL.C., ShiauY.R., ChenP.C., WangH.Y., LaiJ.F., HoM.2002. Vancomycin-resistant enterococci from humans and retail chickens in Taiwan with unique VanB phenotype-vanA genotype incongruence. Antimicrob. Agents Chemother, 46:525–527.
18.
LeclercqR., DerlotE., DuvalJ., CourvalinP.1988. Plasmid-mediated resistance to vancomycin and teicoplanin in Enterococcus faecium. N. Engl. J. Med., 319:157–161.
19.
LeeW.G., HuhJ.Y., ChoS.R., LimY.A.2004. Reduction in glycopeptide resistance in vancomycin-resistant enterococci as a result of vanA cluster rearrangements. Antimicrob. Agents Chemother., 48:1379–1381.
20.
MonteroC.I., StockF., MurrayP.R.2008. Mechanisms of resistance to daptomycin in Enterococcus faecium. Antimicrob. Agents Chemother., 52:1167–1170.
21.
OanceaC., KlareI., WitteW., WernerG.2004. Conjugative transfer of the virulence gene, esp, among isolates of Enterococcus faecium and Enterococcus faecalis. J. Antimicrob. Chemother., 54:232–235.
22.
OhJ.Y., AnS., JinJ.S., LeeY.C., ChoD.T., LeeJ.C.2007. Phenotypic and genotypic differences of the vancomycin-resistant Enterococcus faecium isolates from humans and poultry in Korea. J. Microbiol., 45:466–472.
23.
RiceL.B., CariasL., RudinS., VaelC., GoossensH., KonstabelC., KlareI., NallapareddyS.R., HuangW., MurrayB.E.2003. A potential virulence gene, hylEfm, predominates in Enterococcus faecium of clinical origin. J. Infect. Dis., 187:508–512.
24.
RosenthalV.D., MakiD.G., JamulitratS., MedeirosE.A., TodiS.K, GomezD.Y.et al.2010. International Nosocomial Infection Control Consortium (INICC) report, data summary for 2003–2008, issued June 2009. Am. J. Infect. Control, 38:95–106.
SongJ.H., KoK.S., OhW.S., ParkS., HeoS.T., KwonK.T., RyuS.Y., PeckK.R., LeeN.Y.2006. High frequency of vancomycin-resistant Enterococcus faecium isolates with VanB phenotype and vanA genotype in Korean hospitals. Diagn. Microbiol. Infect. Dis., 56:401–406.
27.
SongJ.H., KoK.S., SuhJ.Y., OhW.S., KangC.I., ChungD.R., PeckK.R., LeeN.Y., LeeW.G.2008. Clinical implications of vancomycin-resistant Enterococcus faecium (VRE) with VanD phenotype and vanA genotype. J. Antimicrob. Chemother., 61:838–844.
28.
TenoverF.C., ArbeitR.D., GoeringR.V., MickelsenP.A., MurrayB.E., PersingD.H., SwaminathanB.1995. Interpreting chromosomal DNA restriction patterns produced by pulsed-field gel electrophoresis:criteria for bacterial strain typing. J. Clin. Microbiol., 33:2233–2239.
29.
TopJ., WillemsR., BlokH., de RegtM., JalinkK., TroelstraA., GoorhuisB., BontenM.2007. Ecological replacement of Enterococcus faecalis by multiresistant clonal complex 17 Enterococcus faecium. Clin. Microbiol. Infect., 13:316–319.
ValdezateS., LabayruC., NavarroA., ManteconM.A., OrtegaM., CoqueT.M., GarcıaM., Saez-NietoJ.A.2009. Large clonal outbreak of multidrug-resistant CC17 ST17 Enterococcus faecium containing Tn5382 in a Spanish hospital. J. Antimicrob. Chemother., 63:17–20.
32.
van SchaikW., TopJ., RileyD.R., BoekhorstJ., VrijenhoekJ.E.P., SchapendonkC.M.E., HendrickxA.P.A., NijmanI.J., BontenM.J.M., TettelinH., WillemsR.J.L.2010. Pyrosequencing-based comparative genome analysis of the nosocomial pathogen Enterococcus faecium and identification of a large transferable pathogenicity island. BMC Genomics, 11:239.
33.
VankerckhovenV., AutgaerdenT.V., VaelC., LammensC., ChapelleS., RossiR., JabesD., GoossensH.2004. Development of multiplex PCR for the detection of asa1, gelE, cylA, esp, and hyl genes in enterococci and survey for virulence determinants among European hospital isolates of Enterococcus faecium. J. Clin. Microbiol., 42:4473–4479.
34.
WernerG., KlareI., WitteW.1997. Arrangement of the vanA gene cluster in enterococci of different ecological origin. FEMS. Microbiol. Lett., 155:55–61.
35.
WillemsR.J., TopJ., van SantenM., RobinsonD.A., CoqueT.M., BaqueroF., GrundmannH., BontenM.J.M.2005. Global spread of vancomycin-resistant Enterococcus faecium from distinct nosocomial genetic complex. Emerg. Infect. Dis., 11:821–828.
36.
WillemsR.J., HomanW., TopJ., van Santen-VerheuvelM., TribeD., ManziorosX.2001. Variant esp gene as a marker of a distinct genetic lineage of vancomycin-resistant Enterococcus faecium spreading in hospitals. Lancet, 357:853–855.
37.
ZhengB., TomitaH., XiaoY.H., WangS., LiY., IkeY.2007. Molecular characterization of vancomycin-resistant Enterococcus faecium isolates from mainland China. J. Clin. Microbiol., 45:2813–2818.
