JAC Advance Access published online on March 10, 2005
Journal of Antimicrobial Chemotherapy, doi:10.1093/jac/dki073
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1 4th Department of Internal Medicine, Molecular Biology Section, University of Athens Medical School, Athens; University General Hospital ‘Attikon’, Rimini 1, 124 62 Chaidari, Greece
* To whom correspondence should be addressed. Objectives: A clinical isolate of Enterobacter cloacae exhibiting reduced susceptibility to imipenem and a positive EDTA-disc synergy test was studied for carbapenemase production. Materials and methods: MICs were determined with standard procedures as well as using a higher inoculum. Isoelectric focusing of cell extracts was used for detection of Results: The isolate was resistant to extended-spectrum Conclusion: We describe a novel class 1 integron containing blaVIM-1 and aac(6')-IIc genes in an E. cloacae clinical isolate.
Received October 15, 2004
Revised January 24, 2005
Accepted January 24, 2005
Original article
Characterization of a new integron containing blaVIM-1 and aac(6')-IIc in an Enterobacter cloacae clinical isolate from Greece
Helen Giamarellou, E-mail: hgiama{at}ath.forthnet.gr
Abstract 
-lactamases. PCR assays with primers specific for the blaVIM gene and the conserved segments of class 1 integrons and sequence analyses were carried out to identify the gene and to map the metallo--lactamase encoding integron. Transferability of the gene was assessed with conjugation experiments using the filter mating technique. To identify the location of the blaVIM-1 gene, Southern hybridization was carried out in genomic DNA using an internal fragment of the blaVIM-1 gene as a probe, amplified by PCR.-lactams. The MICs of carbapenems were below the resistance breakpoints but rose above resistance breakpoints when an inoculum of 108 cfu/mL was used. Isoelectric focusing detected a -lactamase with a pI of 6.1, which exhibited imipenem-hydrolysing activity in a microbiological assay. Ceftazidime and imipenem resistance were not transferable by conjugation. PCR assays identified the blaVIM-1 gene in the variable region of a class 1 integron which also carried the aac(6')-IIc gene. The blaVIM-1 probe hybridized with an approximately 130 kb fragment of genomic DNA, suggesting a chromosomal location of the gene.-lactamases; carbapenemases; E. cloacae.
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