Characterization of pKP1780, a novel IncR plasmid from the emerging Klebsiella pneumoniae ST147, encoding the VIM-1 metallo-b-lactamase

Objectives: To determine the complete nucleotide sequence of the VIM-1-encoding plasmid pKP1780 from Kleb-siella pneumoniae ST147 representing a distinct group of IncR replicons. Methods: The plasmid pKP1780 was from a K. pneumoniae clinical strain (KP-1780) isolated in Greece in 2009. Plasmid DNA was extracted from an Escherichia coli DH5a transformant and sequenced using the 454 Genome Sequencer GS FLX procedure on a standard fragment DNA library. Contig gaps were filled by sequencing of PCR-produced fragments.AnnotationandcomparativeanalysiswereperformedusingsoftwareavailableontheInternet. Results: Plasmid pKP1780 (49770 bp) consisted of an IncR-related sequence (12083 bp) including replication and stability systems, and a multidrug resistance (MDR) mosaic region (37687 bp). blaVIM-1 along with the aacA7, dfrA1 and aadA1 cassettes comprised the variable region of an integron similar to In-e541 from pNL194. The mosaic structure also included the strA, strB, aphA1andmphA resistancegenesaswell as intact (n10) ordefective (n3) insertion sequences and fragments of various transposons. Conclusions: Themosaic structure of pKP1780 exhibitedhigh similaritywith the acquired region of the IncNplasmid pNL194, indicating the acquisition of the VIM-1-encoding MDR region from pNL194 by an IncR-type plasmid