OBJECTIVES: We determined the complete nucleotide sequence of pKOX105, a 54 641 bp plasmid from a Klebsiella oxytoca strain that was isolated from a resident of a long-term-care facility in Bolzano, Italy. METHODS: The plasmid was sequenced using a shotgun approach. Combinatorial PCRs, directed PCRs and walking reads were used to assemble the contigs and to fill in gaps. Gene sequences were compared with reference plasmids and aligned with GenBank data using BLAST and CLUSTAL W software. RESULTS: pKOX105 belonged to incompatibility group IncN, harboured bla(VIM-1), bla(SHV-12), qnrS1, aacA4 and dfrA14 and conferred resistance to carbapenems, oxyimino-cephalosporins, quinolones, aminoglycosides and trimethoprim. It was highly related to the p9 and p12 plasmids from Klebsiella pneumoniae and K. oxytoca strains isolated at a New York City hospital in 2005 carrying bla(KPC-2) and bla(KPC-3), respectively. CONCLUSIONS: IncN plasmids are broad host-range plasmids that have contributed significantly to the worldwide dissemination of many different resistance genes in Enterobacteriaceae from animal and human sources. This plasmid family is now playing a crucial role in the global spread of diverse carbapenemase genes in Klebsiella spp.
OBJECTIVES: We determined the complete nucleotide sequence of pKOX105, a 54 641 bp plasmid from a Klebsiella oxytoca strain that was isolated from a resident of a long-term-care facility in Bolzano, Italy. METHODS: The plasmid was sequenced using a shotgun approach. Combinatorial PCRs, directed PCRs and walking reads were used to assemble the contigs and to fill in gaps. Gene sequences were compared with reference plasmids and aligned with GenBank data using BLAST and CLUSTAL W software. RESULTS: pKOX105 belonged to incompatibility group IncN, harboured bla(VIM-1), bla(SHV-12), qnrS1, aacA4 and dfrA14 and conferred resistance to carbapenems, oxyimino-cephalosporins, quinolones, aminoglycosides and trimethoprim. It was highly related to the p9 and p12 plasmids from Klebsiella pneumoniae and K. oxytoca strains isolated at a New York City hospital in 2005 carrying bla(KPC-2) and bla(KPC-3), respectively. CONCLUSIONS: IncN plasmids are broad host-range plasmids that have contributed significantly to the worldwide dissemination of many different resistance genes in Enterobacteriaceae from animal and human sources. This plasmid family is now playing a crucial role in the global spread of diverse carbapenemase genes in Klebsiella spp.
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