Characterization of the IncA/C plasmid pCC416 encoding VIM-4 and CMY-4 β-lactamases

• Published in: Journal of antimicrobial chemotherapy Vol. 60; no. 2; pp. 258 - 262
• Main Authors: Colinon, Céline, Miriagou, Vivi, Carattoli, Alessandra, Luzzaro, Francesco, Rossolini, Gian Maria
• Format: Journal Article
• Language: English
• Published: Oxford Oxford University Press 01.08.2007; Oxford University Press (OUP)
• Subjects: Antibiotics. Antiinfectious agents. Antiparasitic agents; beta-Lactamases - genetics; Biological and medical sciences; cephalosporinases; Cloning, Molecular; DNA Transposable Elements - genetics; DNA, Bacterial - genetics; Drug Resistance, Multiple, Bacterial - genetics; Enterobacteriaceae - enzymology; Enterobacteriaceae - genetics; Escherichia coli - genetics; Life Sciences; Medical sciences; metallo-β-lactamases; Microbial Sensitivity Tests; Molecular Sequence Data; multiresistance; Pharmacology. Drug treatments; Plasmids - genetics; Polymorphism, Restriction Fragment Length; Reverse Transcriptase Polymerase Chain Reaction; multiresistance; cephalosporinases; metallo-β-lactamases; Plasmid; Enzyme; Multiple resistance; Hydrolases; β-Lactamase; teicoplanin; Bacterial; Escherichia coli; Molecular Sequence Data; Restriction Fragment Length; dalfopristin plus quinupristin; plasmid; salinomycin; Molecular; nitrofurantoin; bacterium isolation; Microbial Sensitivity Tests; transposon; antibiotic resistance; erythromycin; gentamicin; vancomycin; florfenicol; bacterium isolate; bacitracin; polymerase chain reaction; Drug Resistance; Enterobacteriaceae; Multiple; gene transfer; penicillin G; Cloning; vancomycin resistant Enterococcus; kanamycin; bacterial strain; Reverse Transcriptase Polymerase Chain Reaction; article; DNA Transposable Elements; genotype; minimum inhibitory concentration; Enterococcus faecium; Plasmids; replicon; bambermycin; DNA; beta-Lactamases; chloramphenicol; tetracycline; amplicon; nonhuman; streptomycin; Polymorphism; ciprofloxacin
• ISSN: 0305-7453; 1460-2091
• DOI: 10.1093/jac/dkm171

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 β-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.