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International Journal of Medical Microbiology
Volume 298, Issues 5-6, 1 July 2008, Pages 515-527
 
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doi:10.1016/j.ijmm.2007.05.008    How to Cite or Link Using DOI (Opens New Window)
Copyright © 2007 Elsevier GmbH All rights reserved.

Increasing rates of vancomycin resistance among Enterococcus faecium isolated from German hospitals between 2004 and 2006 are due to wide clonal dissemination of vancomycin-resistant enterococci and horizontal spread of vanA clusters

Guido WernerCorresponding Author Contact Information, a, E-mail The Corresponding Author, Ingo Klarea, Carola Fleigea and Wolfgang Wittea

aRobert Koch Institute, Wernigerode Branch, Burgstraße 37, D-38855 Wernigerode, Germany

Received 5 March 2007; 
revised 20 May 2007; 
accepted 24 May 2007. 
Available online 30 October 2007.

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Abstract

Results of national and international surveillance studies revealed increasing rates of vancomycin-resistant Enterococcus faecium (VREF) among German hospital patients since 2003. To investigate the molecular background of vanA-type glycopeptide resistance, 51 clinical VREF isolated between 2004 and 2006 and originating from 19 German hospitals representing 10 Federal States have been investigated. Isolates were characterised by multi-locus sequence typing (MLST), SmaI macrorestriction analysis in pulsed-field gel electrophoresis (PFGE), and multiple-locus variable-number tandem repeat analysis (MLVA). Phylogenetic relatedness between strains was identified using BioNumerics and eBURST software. Distribution of virulence markers esp and hylEfm was investigated by PCR. The structure of the vanA gene clusters was investigated by PCR, long-template PCR, sequencing and Southern hybridisations. The 51 VREF were rather diverse constituting different strain types, different virulence markers and vanA clusters. Within this diversity we found supportive data for a dissemination of related – already vancomycin-resistant – E. faecium among various hospitals and Federal States and for spread of identical vanA gene clusters among clonally different strain types within single hospitals. In conclusion, the increase in the rates of VREF among German hospital patients within the last 2 years might be rather complex and due to different molecular events and scenarios.

Keywords: Vancomycin-resistant enterococci; Vancomycin resistance; MLST; vanA; Enterococcus faecium

Article Outline

Introduction
Materials and methods
Bacterial isolates
Antibiotic susceptibility testing
DNA isolation and PCR
Long-template PCR
DNA sequencing
Assignment of vanA cluster types
Southern-blot, hybridisation and immunological detection
Macrorestriction analysis in PFGE
MLST
MLVA
Results
Antibiotic susceptibility patterns
Typing
PCR for virulence genes
Assignment of Tn1546 types
Discussion
Acknowledgements
References



 
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