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Journal of Clinical Microbiology, August 2000, p. 2885-2888, Vol. 38, No. 8
0095-1137/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

PCR Fragment Length Polymorphism Analysis of Vancomycin-Resistant Enterococcus faecium

Susan Donabedian,1,2,3 Ellie Hershberger,1,2,3 Lee Ann Thal,1,2,3 J. W. Chow,1,2,4 Don B. Clewell,5,6 Barbara Robinson-Dunn,7,8 and Marcus J. Zervos1,2,3,4,9,*

Departments of Medicine,1 Clinical Pathology,9 and Biologic and Materials Sciences5 and Sections of Infectious Disease2 and Microbiology and Disease Surveillance,7 Wayne State University,4 Detroit, William Beaumont Hospital, Royal Oak,3 Michigan Department of Community Health, Lansing,8 and The University of Michigan, Ann Arbor,6 Michigan

Received 7 February 2000/Returned for modification 7 March 2000/Accepted 3 April 2000

In this study, the glycopeptide resistance element, Tn1546, in 124 VanA Enterococcus faecium clinical isolates from 13 Michigan hospitals was evaluated using PCR fragment length polymorphism. There were 26 pulsed-field gel electrophoresis (PFGE) types, which consisted of epidemiologically related and unrelated isolates from separate patients (1992 to 1996). Previously published oligonucleotides specific for regions in the vanA gene cluster of Tn1546 were used to amplify vanRS, vanSH, vanHAX, vanXY, and vanYZ. The glycopeptide resistance element, Tn1546, of E. faecium 228 was used as the basis of comparison for all the isolates in this study. Five PCR fragment length patterns were found, as follows. (i) PCR amplicons were the same size as those of EF228 for all genes in the vanA cluster in 19.4% of isolates. (ii) The PCR amplicon for vanSH was larger than that of EF228 (3.7 versus 2.3 kb) due to an insertion between the vanS and vanH genes (79.2% of isolates). (iii) One isolate in a unique PFGE group had a vanSH amplicon larger than that of EF228 (5.7 versus 2.3 kb) due to an insertion in the vanS gene and an insertion between the vanS and vanH genes. (iv) One isolate did not produce a vanSH amplicon, but when vanS and vanH were amplified separately, both amplicons were the same size as those as EF228. (v) One isolate had a vanYZ PCR product larger than that of EF228 (2.8 versus 1.6 kb). This study shows that in a majority of the VanA E. faecium isolates, Tn1546 is altered compared to that of EF228. A total of 79.2% of the study isolates had the same-size insertion between the vanS and vanH genes. The results of this study show dissemination of an altered Tn1546 in heterologous VanA E. faecium in Michigan hospitals.

* Corresponding author. Mailing address: William Beaumont Hospital, Research Institute, 3601 West 13 Mile Rd., Royal Oak, MI 48073. Phone: (248) 551-0419. Fax: (248) 551-5069. E-mail: MZervos{at}Beaumont.edu.

Journal of Clinical Microbiology, August 2000, p. 2885-2888, Vol. 38, No. 8
0095-1137/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


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