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Molecular characterisation of linezolid resistance in two vancomycin-resistant (VanB) Enterococcus faecium isolates using Pyrosequencing™

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Abstract

In this paper, we describe the phenotypic and molecular characteristics of two clinically relevant, vancomycin-resistant (VanB), linezolid-resistant Enterococcus faecium isolates. Pyrosequencing™ showed the G to T single nucleotide polymorphism at bp 2576 in the genes coding for 23S rRNA and was used to quantify the proportion of G to T mutations among six different 23S rRNA genes in E. faecium as a marker for the molecular level of resistance to linezolid. In both isolates, the G to T mutation was found in two of six alleles, and no further mutations in the genes coding for 23S rRNA were found. The dynamic process of linezolid resistance could be demonstrated by the complete reversion of resistant alleles back to only wild type alleles in consecutive isolates of one isolate. Pyrosequencing™ being used to detect and quantify resistance to linezolid has been proven as a fast and reliable molecular screening method for monitoring linezolid resistance.

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Acknowledgements

We thank Elke Halle, Institute for Microbiology and Hygiene, Charite Medical University, Berlin, Germany, for kindly providing the reference strains of linezolid-resistant enterococci.

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Author notes

  1. M. A. Horstkotte

    Present address: Bioscientia Institut für Medizinische Diagnostik GmbH, Bioscientia MVZ Hamburg, Papenreye 63, 22453, Hamburg, Germany

Authors and Affiliations

  1. Department of Pharmaceutical Biology and Microbiology, University of Hamburg, Bundesstrasse 45, 20146, Hamburg, Germany

    B. Saager, B. S. Timmerbeil & P. Heisig

  2. Department of Medical Microbiology, Virology and Hygiene, University Medical Center Hamburg-Eppendorf, Martinistrasse 52, 20246, Hamburg, Germany

    H. Rohde, G. Franke, S. Scherpe, I. Sobottka & M. A. Horstkotte

  3. Department of Intensive Care Medicine, University Medical Center Hamburg-Eppendorf, Martinistrasse 52, 20246, Hamburg, Germany

    W. Pothmann

  4. Department of Bone Marrow Transplantation, University Medical Center Hamburg-Eppendorf, Martinistrasse 52, 20246, Hamburg, Germany

    J. Dahlke

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  1. B. Saager
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Correspondence to M. A. Horstkotte.

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All authors have seen and approved the final version of the manuscript. There were no conflicts of interests to declare. The experiments comply with the current laws of the country in which they are performed.

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Saager, B., Rohde, H., Timmerbeil, B.S. et al. Molecular characterisation of linezolid resistance in two vancomycin-resistant (VanB) Enterococcus faecium isolates using Pyrosequencing™. Eur J Clin Microbiol Infect Dis 27, 873–878 (2008). https://doi.org/10.1007/s10096-008-0514-6

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  • DOI: https://doi.org/10.1007/s10096-008-0514-6

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