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Journal of Clinical Microbiology, December 2003, p. 5407-5413, Vol. 41, No. 12
0095-1137/03/$08.00+0     DOI: 10.1128/JCM.41.12.5407-5413.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

PCR Typing of Genetic Determinants for Metallo-ß-Lactamases and Integrases Carried by Gram-Negative Bacteria Isolated in Japan, with Focus on the Class 3 Integron

Naohiro Shibata,* Yohei Doi, Kunikazu Yamane, Tetsuya Yagi, Hiroshi Kurokawa, Keigo Shibayama, Haru Kato, Kumiko Kai, and Yoshichika Arakawa

Department of Bacterial Pathogenesis and Infection Control, National Institute of Infectious Diseases, Tokyo, Japan

Received 18 June 2003/ Returned for modification 20 August 2003/ Accepted 27 August 2003

From January 2001 to December 2002, 587 strains of gram-negative bacterial isolates demonstrating resistance to ceftazidime and a combination of sulbactam and cefoperazone were subjected to a disk diffusion screening test using sodium mercaptoacetic acid; 431 strains (73.4%) appeared to produce metallo-ß-lactamase (MBL). Of these 431 strains, 357 were found by PCR to carry genes for IMP-1 type MBL (blaIMP-1), while only 7 and 67 strains carried the IMP-2 gene (blaIMP-2) and the VIM-2 gene (blaVIM-2), respectively. Neither VIM-1 nor SPM-1 type MBL genes were found among the strains tested. Of 431 strains, 427 carried the intI1 gene, and 4 strains carrying both the intI1 and intI3 genes were reidentified as Pseudomonas putida harboring blaIMP-1. Of these four P. putida strains, three strains and one strain, respectively, were separately isolated from two hospitals located in the same prefecture, and the three strains showed very similar pulsed-field gel electrophoresis patterns. Of 357 blaIMP-1 carriers, 116, 53, 51, 47, and 30 strains were identified as Pseudomonas aeruginosa, Alcaligenes xylosoxidans, P. putida/fluorescens, Serratia marcescens, and Acinetobacter baumannii, respectively. Four strains carrying blaIMP-2 were reidentified as P. putida. Sixty-three P. aeruginosa strains and four P. putida strains carried blaVIM-2. Of 427 intI1-positive strains, 180, 53, 51, 47, and 35 were identified as P. aeruginosa, A. xylosoxidans, P. putida/fluorescens, S. marcescens, and A. baumannii, respectively. In the present study, it was confirmed that strains carrying blaIMP-1 with a class 1 integron are the most prevalent type in Japan, although several intI3 carriers have also been identified sporadically in this country.

* Corresponding author. Mailing address: Department of Bacterial Pathogenesis and Infection Control, 4-7-1 Musashi-Murayama, Tokyo 208-0011, Japan. Phone: 81-42-561-0771, ext. 363. Fax: 81-42-561-7173. E-mail: nsbt{at}nih.go.jp.

Journal of Clinical Microbiology, December 2003, p. 5407-5413, Vol. 41, No. 12
0095-1137/03/$08.00+0     DOI: 10.1128/JCM.41.12.5407-5413.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



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