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Journal of Clinical Microbiology, September 2002, p. 3326-3333, Vol. 40, No. 9
0095-1137/02/$04.00+0     DOI: 10.1128/JCM.40.9.3326-3333.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Possible Connection between a Widely Disseminated Conjugative Gentamicin Resistance (pMG1-Like) Plasmid and the Emergence of Vancomycin Resistance in Enterococcus faecium

Haruyoshi Tomita,1 Carl Pierson,2 Suk Kyung Lim,1,{dagger} Don B. Clewell,3,4 and Yasuyoshi Ike1,5*

Department of Microbiology,1 Laboratory of Bacterial Drug Resistance, Gunma University School of Medicine, Maebashi, Gunma 371-8511, Japan,5 Division of Microbiology, Clinical Laboratory, Medical School Hospital,2 Department of Microbiology/Immunology, School of Medicine,4 Department of Biologic and Materials Sciences, School of Dentistry, University of Michigan, Ann Arbor, Michigan 481093

Received 4 February 2002/ Returned for modification 21 April 2002/ Accepted 22 June 2002

A total of 640 vancomycin-resistant Enterococcus faecium (VRE) isolates, which were obtained between 1994 and 1999 from the Medical School Hospital of the University of Michigan, Ann Arbor, were used in this study. Of the 640 strains, 611 and 29 were VanA and VanB VRE, respectively, based on PCR analysis. Four hundred ninety-two (77%) of the strains exhibited resistance to concentrations of gentamicin from 64 µg/ml (MIC) to more than 1,024 µg/ml (MIC). The gentamicin resistance of each of 261 (53%) of the 492 gentamicin-resistant strains was transferred to E. faecium at a frequency of about 10-5 to 10-6 per donor cell in broth mating. More than 90% of vancomycin resistances of the 261 strains cotransferred with the gentamicin resistances to E. faecium strains by filter mating. The conjugative gentamicin resistance plasmids were identified and were classified into five types (A through E) with respect to their EcoRI restriction profiles. The transfer frequencies of each type of plasmid between E. faecium strains or Enterococcus faecalis strains were around 10-3 to 10-5 per donor cell or around 10-6 to 10-7 per donor cell, respectively, in broth mating. Type A and type B were the most frequently isolated, at an isolation frequency of about 40% per VRE isolate harboring the gentamicin resistance conjugative plasmid. The plasmids did not show any homology in Southern hybridization with the pheromone-responsive plasmids and broad-host-range plasmids pAMß1 and pIP501. The EcoRI or NdeI restriction fragments of each type of plasmids hybridized to the conjugative gentamicin resistance plasmid pMG1 (65.1 kb), which was originally isolated from an E. faecium clinical isolate, and transfer efficiently in broth mating.

* Corresponding author. Mailing address: Department of Microbiology, Gunma University School of Medicine, Showa-machi 3-39-22, Maebashi, Gunma 371-8511, Japan. Phone: 81-27-220-7990. Fax: 81-27-220-7996. E-mail: yasuike{at}med.gunma-u.ac.jp.

{dagger} Present address: National Veterinary Research and Quarantine Service, Manan-gu, Anyang kyonggido, Republic of Korea, 430-016.

Journal of Clinical Microbiology, September 2002, p. 3326-3333, Vol. 40, No. 9
0095-1137/02/$04.00+0     DOI: 10.1128/JCM.40.9.3326-3333.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



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Copyright © 2002 by the American Society for Microbiology. All rights reserved.