JAC Advance Access originally published online on May 31, 2007
Journal of Antimicrobial Chemotherapy 2007 60(2):258-262; doi:10.1093/jac/dkm171
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Characterization of the IncA/C plasmid pCC416 encoding VIM-4 and CMY-4 ß-lactamases
1 Dipartimento di Biologia Molecolare, Laboratorio di Fisiologia e Biotecnologia dei Microorganismi, Università di Siena, Siena, Italy 2 Laboratory of Bacteriology, Hellenic Pasteur Institute, Athens, Greece 3 Dipartimento di Malattie Infettive, Istituto Superiore di Sanita, Rome, Italy 4 Laboratorio di Microbiologia, Ospedale di Circolo, Varese, Italy
Received 18 December 2006; returned 5 February 2007; revised 10 March 2007; accepted 24 April 2007
* Corresponding author. Tel: +30-210-6478810; Fax: +30-210-6423498; E-mail: miriagou{at}mail.pasteur.gr
Objectives: To characterize the antibiotic resistance regions of pCC416, a VIM-4- and CMY-4-encoding plasmid from clinical enterobacteria, and to elucidate its relation with the CMY-encoding plasmids widely diffused in Salmonella.
Methods: The enterobacterial multiresistant plasmid pCC416 was derived from an Escherichia coli transconjugant and characterized. Conventional and long-range PCR assays were performed using primers specific for VIM-4- and CMY-4-encoding segments of pCC416. Amplicons were characterized by sequencing. blaVIM-4, blaCMY-4 and IntI1-specific probes were prepared from PCR products and used for the identification of various pCC416 clones. VIM- and CMY-positive BamHI and Sau3AI fragments of pCC416 were cloned into pACYC184 and their sequences were determined by gene walking.
Results: The pCC416 plasmid contained two distinct resistant loci carrying ß-lactamase genes. The blaVIM-4 gene was part of an integron located in a complex, multidrug-resistant region of novel structure, interspersed with mobile elements or remnants thereof and being similar to various regions of other resistance plasmids. Nevertheless, a region in the 3' end of this structure resembled the respective region found in a CMY-2-encoding plasmid from Salmonella. The blaCMY-4 gene was identified within an 11.3 kb region also related to the CMY-2-encoding plasmids.
Conclusions: pCC416 probably evolved from an IncA/C2, CMY-encoding plasmid through acquisition of a VIM-encoding In4-type integron providing an example of accretion of resistance determinants in a single replicon.
Keywords: metallo-ß-lactamases , cephalosporinases , multiresistance
Present address. Laboratoire de Radioécologie et d'Ecotoxicologie, Institut de Radioprotection et de Sûreté nucléaire, DEI/SECRE/LRE, Bât 186, BP 3, F-13115 Saint Paul Lez Durance Cedex, France.
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