1887

Abstract

The aim of this study was to characterize the resistance mechanism in four clinical and five intestinal vancomycin-resistant strains with VanB phenotype recovered from unrelated patients confined in two Spanish hospitals and to determine their clonal relationships. MIC values for vancomycin and teicoplanin were 16–32 and 0.5 μg ml, respectively. The mechanism of vancomycin resistance, as well as the genetic environment of the implicated gene, was analysed by PCR and sequencing. The gene was detected in all nine strains and the intergenic region showed the characteristic mutations of the subtype. Two possibly related PFGE patterns, A (seven strains) and B (two strains), were distinguished among these enterococci. The intergenic region was amplified in the nine strains and two amino acid changes were detected in the protein encoded by the gene in strains of pattern A with respect to those of pattern B. The gene cluster was integrated into Tn in all nine strains, being gene-linked to this transposon. The (6′)-, (3′)- and (B) genes were also detected in all of the strains. Both isolates with PFGE pattern B contained the gene. In summary, -containing strains with indistinguishable PFGE patterns were recovered from seven patients from two Spanish hospitals.

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2006-09-01
2019-10-21
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References

  1. Aarestrup, F. M., Agerso, Y., Gerner-Smidt, P., Madsen, M. & Jensen, L. B. ( 2000; ). Comparison of antimicrobial resistance phenotypes and resistance genes in Enterococcus faecalis and Enterococcus faecium from humans in the community, broilers, and pigs in Denmark. Diagn Microbiol Infect Dis 37, 127–137.[CrossRef]
    [Google Scholar]
  2. Carias, L. L., Rudin, S. D., Donskey, C. J. & Rice, L. B. ( 1998; ). Genetic linkage and cotransfer of a novel vanB-containing transposon (Tn5382) and a low-affinity penicillin-binding protein 5 gene in a clinical vancomycin-resistant Enterococcus faecium isolate. J Bacteriol 180, 4426–4434.
    [Google Scholar]
  3. Cetinkaya, Y., Falk, P. & Mayhall, C. G. ( 2000; ). Vancomycin-resistant enterococci. Clin Microbiol Rev 13, 686–707.[CrossRef]
    [Google Scholar]
  4. Clark, N. C., Olsvik, Ø., Swenson, J. M., Spiegel, C. A. & Tenover, F. C. ( 1999; ). Detection of a streptomycin/spectinomycin adenylyltransferase gene (aadA) in Enterococcus faecalis. Antimicrob Agents Chemother 43, 157–160.[CrossRef]
    [Google Scholar]
  5. Dahl, K. H., Simonsen, G. S., Olsvik, Ø. & Sundsfjord, A. ( 1999; ). Heterogeneity in the vanB gene cluster of genomically diverse clinical strains of vancomycin-resistant enterococci. Antimicrob Agents Chemother 43, 1105–1110.
    [Google Scholar]
  6. Dahl, K. H., Lundblad, E. W., Rokenes, T. P., Olsvik, Ø. & Sundsfjord, A. ( 2000; ). Genetic linkage of the vanB2 gene cluster to Tn5382 in vancomycin-resistant enterococci and characterization of two novel insertion sequences. Microbiology 146, 1469–1479.
    [Google Scholar]
  7. Domingo, M.-C., Huletsky, A., Bernal, A., Giroux, R., Boudreau, D. K., Picard, F. J. & Bergeron, M. G. ( 2005; ). Characterization of a Tn5382-like transposon containing the vanB2 gene cluster in a Clostridium strain isolated from human faeces. J Antimicrob Chemother 55, 466–474.[CrossRef]
    [Google Scholar]
  8. Dunny, G. M., Craig, R. A., Carron, R. L. & Clewell, D. B. ( 1979; ). Plasmid transfer in Streptococcus faecalis: production of multiple sex pheromones by recipients. Plasmid 2, 454–465.[CrossRef]
    [Google Scholar]
  9. Dutka-Malen, S., Evers, S. & Courvalin, P. ( 1995; ). Detection of glycopeptide resistance genotypes and identification to the species level of clinically relevant enterococci by PCR. J Clin Microbiol 33, 24–27.
    [Google Scholar]
  10. Garnier, F., Taourit, S., Glaser, P., Courvalin, P. & Galimand, M. ( 2000; ). Characterization of transposon Tn1549, conferring VanB-type resistance in Enterococcus spp. Microbiology 146, 1481–1489.
    [Google Scholar]
  11. Gholizadeh, Y. & Courvalin, P. ( 2000; ). Acquired and intrinsic glycopeptide resistance in enterococci. Int J Antimicrob Agents 16, S11–S17.[CrossRef]
    [Google Scholar]
  12. Gold, H. S., Ünal, S., Cercenado, E., Thauvin-Eliopoulos, C., Eliopoulos, G. M., Wennersten, C. B. & Moellering, R. C., Jr ( 1993; ). A gene conferring resistance to vancomycin but not teicoplanin in isolates of Enterococcus faecalis and Enterococcus faecium demonstrates homology with vanB, vanA, and vanC genes of enterococci. Antimicrob Agents Chemother 37, 1604–1609.[CrossRef]
    [Google Scholar]
  13. Granlund, M., Carlsson, C., Edebro, H., Emanuelsson, K. & Lundholm, R. ( 2006; ). Nosocomial outbreak of vanB2 vancomycin-resistant Enterococcus faecium in Sweden. J Hosp Infect 62, 254–256.[CrossRef]
    [Google Scholar]
  14. Jureen, R., Top, J., Mohn, S. C., Harthug, S., Langeland, N. & Willens, R. J. L. ( 2003; ). Molecular characterization of ampicillin-resistant Enterococcus faecium isolates from hospitalized patients in Norway. J Clin Microbiol 41, 2330–2336.[CrossRef]
    [Google Scholar]
  15. Kawalec, M., Gniadkowski, M., Zielinska, U., Klos, W. & Hryniewicz, W. ( 2001; ). Vancomycin-resistant Enterococcus faecium strain carrying the vanB2 gene variant in a Polish hospital. J Clin Microbiol 39, 811–815.[CrossRef]
    [Google Scholar]
  16. Klare, I., Konstabel, C., Mueller-Bertling, S. & 10 other authors ( 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.[CrossRef]
    [Google Scholar]
  17. Lee, W. G., Jernigan, J. A., Rashed, J. K., Anderson, G. J. & Tenover, F. C. ( 2001; ). Possible horizontal transfer of the vanB2 gene among genetically diverse strains of vancomycin-resistant Enterococcus faecium in a Korean hospital. J Clin Microbiol 39, 1165–1168.[CrossRef]
    [Google Scholar]
  18. Lorenzo-Díaz, F., Delgado, T., Reyes-Darias, J. A., Flores, C., Méndez-Álvarez, S., Villar, J., Sierra, A. & Claverie-Martín, F. ( 2004; ). Characterization of the first VanB vancomycin-resistant Enterococcus faecium isolated in a Spanish hospital. Curr Microbiol 48, 199–203.[CrossRef]
    [Google Scholar]
  19. Lu, J.-J., Perng, C.-L., Ho, M.-F., Chiueh, T.-S. & Lee, W.-H. ( 2001; ). High prevalence of VanB2 vancomycin-resistant Enterococcus faecium in Taiwan. J Clin Microbiol 39, 2140–2145.[CrossRef]
    [Google Scholar]
  20. Lu, J.-J., Chang, T.-Y., Perng, C.-L. & Lee, S.-Y. ( 2005; ). The vanB2 gene cluster of the majority of vancomycin-resistant Enteroccus faecium isolates from Taiwan is associated with the pbp5 gene and is carried by Tn5382 containing a novel insertion sequence. Antimicrob Agents Chemother 49, 3937–3939.[CrossRef]
    [Google Scholar]
  21. Mato, R., de Lencastre, H., Roberts, R. B. & Tomasz, A. ( 1996; ). Multiplicity of genetic backgrounds among vancomycin-resistant Enterococcus faecium isolates recovered from an outbreak in a New York City hospital. Microb Drug Resist 2, 309–317.[CrossRef]
    [Google Scholar]
  22. McGregor, K. F. & Young, H.-K. ( 2000; ). Identification and characterization of vanB2 glycopeptide resistance elements in enterococci isolated in Scotland. Antimicrob Agents Chemother 44, 2341–2348.[CrossRef]
    [Google Scholar]
  23. McGregor, K. F., Nolan, C., Young, H.-K., Palepou, M.-F. I., Tysall, L. & Woodford, N. ( 2001; ). Prevalence of the vanB2 gene cluster in the VanB glycopeptide-resistant enterococci in the United Kingdom and the Republic of Ireland and its association with a Tn5382-like element. Antimicrob Agents Chemother 45, 367–368.[CrossRef]
    [Google Scholar]
  24. Miele, A., Bandera, M. & Goldstein, B. P. ( 1995; ). Use of primers selective for vancomycin resistance genes to determine van genotype in enterococci and to study gene organization in VanA isolates. Antimicrob Agents Chemother 39, 1772–1778.[CrossRef]
    [Google Scholar]
  25. Murray, B. ( 1997; ). Vancomycin-resistant enterococci. Am J Med 102, 284–293.[CrossRef]
    [Google Scholar]
  26. NCCLS ( 2002; ). Methods for dilution antimicrobial susceptibility tests for bacteria that grow aerobically. NCCLS document M7-A5. Wayne, PA: National Committee for Clinical Laboratory Standards.
  27. Perichon, B., Reynolds, P. & Courvalin, P. ( 1997; ). VanD-type glycopeptide-resistant Enterococcus faecium BM4339. Antimicrob Agents Chemother 41, 2016–2018.
    [Google Scholar]
  28. Quintiliani, R., Jr, Evers, S. & Courvalin, P. ( 1993; ). The vanB gene confers various levels of self-transferable resistance to vancomycin in enterococci. J Infect Dis 167, 1220–1223.[CrossRef]
    [Google Scholar]
  29. Rice, L. B., Bellais, S., Carias, L. L., Hutton-Thomas, R., Bonomo, R. A., Caspers, P., Page, M. G. P. & Gutmann, L. ( 2004; ). Impact of specific pbp5 mutations on expression of β-lactam resistance in Enterococcus faecium. Antimicrob Agents Chemother 48, 3028–3032.[CrossRef]
    [Google Scholar]
  30. Simonsen, G. S., Andersen, B. M., Digranes, A. & 9 other authors ( 1998; ). Low faecal carrier rate of vancomycin resistant enterococci in Norwegian hospital patients. Scand J Infect Dis 30, 465–468.[CrossRef]
    [Google Scholar]
  31. Sutcliffe, J., Grebe, T., Tait-Kamradt, A. & Wondrack, L. ( 1996; ). Detection of erythromycin-resistant determinants by PCR. Antimicrob Agents Chemother 40, 2562–2566.
    [Google Scholar]
  32. Tenover, F. C., Arbeit, R. D., Goering, R. V., Mickelsen, P. A., Murray, B. E., Persing, D. H. & Swaminathan, B. ( 1995; ). Interpreting chromosomal DNA restriction patterns produced by pulsed-field gel electrophoresis: criteria for bacterial strain typing. J Clin Microbiol 33, 2233–2239.
    [Google Scholar]
  33. Torres, C., Reguera, J. A., Sanmartín, M. J., Pérez-Díaz, J. C. & Baquero, F. ( 1994; ). vanA-mediated vancomycin-resistant Enterococcus spp. in sewage. J Antimicrob Chemother 33, 553–561.[CrossRef]
    [Google Scholar]
  34. Torres, C., Tenorio, C., Portillo, A., García, M., Martínez, C., del Campo, R., Ruiz-Larrea, F. & Zarazaga, M. ( 2003; ). Intestinal colonization by vanA- or vanB2-containing enterococcal isolates of healthy animals in Spain. Microb Drug Resist 9 (Suppl. 1), S47–S52.
    [Google Scholar]
  35. Turabelidze, D., Kotetishvili, M., Krefer, A., Morris, J. G. & Sulakvelidze, A. ( 2000; ). Improved pulsed-field gel electrophoresis for typing vancomycin-resistant enterococci. J Clin Microbiol 38, 4242–4245.
    [Google Scholar]
  36. Umeda, A., Garnier, F., Courvalin, P. & Galimand, M. ( 2002; ). Association between the vanB2 glycopeptide resistance operon and Tn1549 in enterococci from France. J Antimicrob Chemother 50, 253–256.[CrossRef]
    [Google Scholar]
  37. Van de Klundert, J. A. M. & Vliegenthart, J. S. ( 1993; ). PCR detection of genes for aminoglycoside-modifying enzymes. In Diagnostic Molecular Microbiology: Principles and Applications, pp. 547–552. Edited by D. H. Persing, T. F. Smith, F. C. Tenover & T. J. White. Washington, DC: American Society for Microbiology.
  38. Woodford, N. ( 2001; ). Epidemiology of the genetic elements responsible for acquired glycopeptide resistance in enterococci. Microb Drug Resist 7, 229–236.[CrossRef]
    [Google Scholar]
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