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Antimicrobial Agents and Chemotherapy, May 1998, p. 1229-1232, Vol. 42, No. 5
0066-4804/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.

A New High-Level Gentamicin Resistance Gene, aph(2")-Id, in Enterococcus spp.

Shane F. Tsai,1 Marcus J. Zervos,2 Don B. Clewell,3 Susan M. Donabedian,2 Daniel F. Sahm,4,dagger and Joseph W. Chow1,5,*

Research and Medical Service, Department of Veterans' Affairs Medical Center,1 and Wayne State University School of Medicine,5 Detroit, Michigan 48201; William Beaumont Hospital, Royal Oak, Michigan 480732; Departments of Biologic and Materials Sciences and Microbiology and Immunology, University of Michigan, Ann Arbor, Michigan 481093; and Department of Pathology, Jewish Hospital at Washington University Medical Center, St. Louis, Missouri 631104

Received 5 August 1997/Returned for modification 3 December 1997/Accepted 18 February 1998

Enterococcus casseliflavus UC73 is a clinical blood isolate with high-level resistance to gentamicin. DNA preparations from UC73 failed to hybridize with intragenic probes for aac(6')-Ie-aph(2")-Ia and aph(2")-Ic. A 4-kb fragment from UC73 was cloned and found to confer resistance to gentamicin in Escherichia coli DH5alpha transformants. Nucleotide sequence analysis revealed the presence of a 906-bp open reading frame whose deduced amino acid sequence had a region with homology to the aminoglycoside-modifying enzyme APH(2")-Ic and to the C-terminal domain of the bifunctional enzyme AAC(6')-APH(2"). The gene is designated aph(2")-Id, and its observed phosphotransferase activity is designated APH(2")-Id. A PCR-generated intragenic probe hybridized to the genomic DNA from 17 of 118 enterococcal clinical isolates (108 with high-level gentamicin resistance) from five hospitals. All 17 were vancomycin-resistant Enterococcus faecium isolates, and pulsed-field typing revealed three distinct clones. The combination of ampicillin plus either amikacin or neomycin exhibited synergistic killing against E. casseliflavus UC73. Screening and interpretation of high-level aminoglycoside resistance in enterococci may need to be modified to include detection of APH(2")-Id.

* Corresponding author. Mailing address: Division of Infectious Diseases, Wayne State University School of Medicine, 4160 John R, Suite 2140, Detroit, MI 48201-2021. Phone: (313) 745-9649. Fax: (313) 763-9905. E-mail: waichung{at}umich.edu.

dagger Present address: MRL Pharmaceutical Services, Inc., Reston, VA 20190.

Antimicrobial Agents and Chemotherapy, May 1998, p. 1229-1232, Vol. 42, No. 5
0066-4804/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.


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