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Antimicrobial Agents and Chemotherapy, May 1999, p. 1105-1110, Vol. 43, No. 5
0066-4804/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.
Heterogeneity in the vanB Gene Cluster of Genomically
Diverse Clinical Strains of Vancomycin-Resistant Enterococci
Kristin Hegstad
Dahl,*
Gunnar Skov
Simonsen,
Ørjan
Olsvik, and
Arnfinn
Sundsfjord
Department of Medical Microbiology,
University and University Hospital of Tromsø, Tromsø, Norway
Received 3 November 1998/Returned for modification 7 January
1999/Accepted 3 March 1999
Molecular analysis of 17 genomically unrelated clinical VanB-type
vancomycin-resistant enterococcus isolates from hospital patients in
Germany, Norway, Sweden, the United Kingdom and the United States
revealed three subtypes of the vanB gene
cluster
vanB1, vanB2, and
vanB3
which was in accordance with previous subtyping of
the ligase gene sequence. There was no correlation between vanB subtype and levels of vancomycin resistance. All
strains studied carried a structurally conserved vanB gene
cluster as shown by long-range PCR (long PCR) covering 5,959 bp of the
published sequence in vanB1 strain V583. Restriction
analysis of long PCR amplicons displayed one unique vanB1
pattern and a second vanB2- and vanB3-specific
pattern. The vanSB-vanYB intergenic
sequences with flanking coding regions were identical within each
vanB subtype with one exception. A U.S. vanB2
isolate had a 789-bp enlargement of this region containing a putative
open reading frame (ORF) with substantial homology to an ORF in the
Clostridium perfringens IS1469 insertion
element. The molecular heterogeneity within the vanB gene
cluster has implications for the selection of PCR primers, as the
primers must ensure detection of all vanB subtypes, and is
of importance when considering reservoirs and dissemination of
vanB resistance. The molecular identity within the
vanB1 and the vanB2 subtype indicates
horizontal transmission of both gene clusters between isolates in
different geographical areas. Restriction analysis of long PCR
vanB amplicons may reveal specific varieties that can be
used as epidemiological markers for mobile determinants conferring
VanB-type resistance. The finding of three distinct vanB
gene clusters should encourage a search for different environmental reservoirs of vanB resistance determinants.
*
Corresponding author. Mailing address: Department of
Medical Microbiology, University of Tromsø, N-9037 Tromsø, Norway.
Phone: 47 77 64 57 62. Fax: 47 77 64 53 50. E-mail:
kristind{at}fagmed.uit.no.
Antimicrobial Agents and Chemotherapy, May 1999, p. 1105-1110, Vol. 43, No. 5
0066-4804/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.
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