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Journal of Clinical Microbiology, September 2001, p. 3115-3121, Vol. 39, No. 9
0095-1137/01/$04.00+0   DOI: 10.1128/JCM.39.9.3115-3121.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Identification of Aminoglycoside-Modifying Enzymes by Susceptibility Testing: Epidemiology of Methicillin-Resistant Staphylococcus aureus in Japan

Takashi Ida,1 Ryoichi Okamoto,1 Chieko Shimauchi,1 Toyoji Okubo,2 Akio Kuga,1 and Matsuhisa Inoue1,*

Department of Microbiology, Kitasato University School of Medicine, 1-15-1 Kitasato, Sagamihara, Kanagawa 228-8555,1 and Laboratory of Drug Resistance in Bacteria, Gunma University School of Medicine, 3-39-22 Showamachi, Maebashi, Gunma 371-8511,2 Japan

Received 29 January 2001/Returned for modification 31 March 2001/Accepted 15 June 2001

A multiple-primer PCR was used to identify genes encoding aminoglycoside-modifying enzymes in 381 clinical isolates of methicillin-resistant Staphylococcus aureus (MRSA). The technique used three sets of primers delineating specific DNA fragments of the aph(3')-III, ant(4')-I, and aac(6')-aph(2") genes, which influence the MICs of gentamicin, tobramycin, and lividomycin. Isolates with none of the three genes detected were susceptible to all three agents. Isolates with the aph(3')-III gene showed resistance to lividomycin (MIC > 1,024 µg/ml), and those with the ant(4')-I gene were resistant to tobramycin (MIC >=  8 µg/ml). Isolates with only the aac(6')-aph(2") gene were resistant to gentamicin (MIC >=  8 µg/ml) and tobramycin in decreasing order; those with both the ant(4')-I and aac(6')-aph(2") genes also were resistant to gentamicin and tobramycin, but in increasing order. Susceptibility testing, then, could detect specific genes. In 381 Japanese MRSA isolates, the ant(4')-I, aac(6')-aph(2"), and aph(3')-III genes were prevalent in 84.5, 61.7, and 8.9%, respectively. Isolates with only the ant(4')-I gene had coagulase type II or III, but isolates with both the ant(4')-I and aac(6')-aph(2") genes included all coagulase types. Most isolates with coagulase type IV or VII carried the aac(6')-aph(2") gene. Of the MRSA isolates with ant(4')-I and/or aac(6')-aph(2") genes, 97% were resistant to aminoglycosides in clinical use, but a new aminoglycoside, arbekacin, had excellent activity against these isolates.

* Corresponding author. Mailing address: Department of Microbiology, Kitasato University School of Medicine, 1-15-1 Kitasato, Sagamihara, Kanagawa 228-8555, Japan. Phone: 81-42-778-9355. Fax: 81-42-778-9350. E-mail: matsu{at}kitasato-u.ac.jp.

Journal of Clinical Microbiology, September 2001, p. 3115-3121, Vol. 39, No. 9
0095-1137/01/$04.00+0   DOI: 10.1128/JCM.39.9.3115-3121.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


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