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

Abstract

ABSTRACT 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-β-lactamase (MBL). Of these 431 strains, 357 were found by PCR to carry genes for IMP-1 type MBL ( bla IMP-1 ), while only 7 and 67 strains carried the IMP-2 gene ( bla IMP-2 ) and the VIM-2 gene ( bla VIM-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 bla IMP-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 bla IMP-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 bla IMP-2 were reidentified as P. putida . Sixty-three P. aeruginosa strains and four P. putida strains carried bla VIM-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 bla IMP-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.