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Antimicrobial Agents and Chemotherapy, February 2006, p. 534-541, Vol. 50, No. 2
0066-4804/06/$08.00+0     doi:10.1128/AAC.50.2.534-541.2006
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

Horizontal Transfer of bla_CMY-Bearing Plasmids among Clinical Escherichia coli and Klebsiella pneumoniae Isolates and Emergence of Cefepime-Hydrolyzing CMY-19

Department of Bacterial Pathogenesis and Infection Control, National Institute of Infectious Diseases, Tokyo,¹ Department of Bacteriology and Bacterial Infection Control, Gunma University Graduate School of Medicine, Gunma, Japan²

Received 30 June 2005/ Returned for modification 17 August 2005/ Accepted 4 November 2005

Nine Escherichia coli and 5 Klebsiella pneumoniae clinical isolates resistant to various cephalosporins and cephamycins were identified in a Japanese general hospital between 1995 and 1997. All nine E. coli isolates and one K. pneumoniae isolate carried bla_CMY-9, while the other four K. pneumoniae isolates harbored a variant of bla_CMY-9, namely, bla_CMY-19. The pulsed-field gel electrophoresis patterns of the nine CMY-9-producing E. coli isolates were almost identical, suggesting their clonal relatedness, while those of the five K. pneumoniae isolates were divergent. Plasmid profiles, Southern hybridization, and conjugation assays revealed that the genes for the CMY-9 and the CMY-19 ß-lactamases were located on very similar conjugative plasmids in E. coli and K. pneumoniae. The genetic environment of bla_CMY-19 was identical to that of bla_CMY-9. A single amino acid substitution, I292S, adjacent to the H-10 helix region was observed between CMY-9 and CMY-19. This substitution was suggested to be responsible for the expansion of the hydrolyzing activity against several broad-spectrum cephalosporins, and this finding was consistent with the kinetic parameters determined with purified enzymes. These findings suggest that the bla_CMY-19 genes found in the four K. pneumoniae isolates might have originated from bla_CMY-9 gene following a point mutation and dispersed among genetically different K. pneumoniae isolates via a large transferable plasmid.

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