title: Whole-genome assembly of Klebsiella pneumoniae coproducing NDM-1 and OXA-232 carbapenemases using single-molecule, real-time sequencing.
abstract: The whole-genome sequence of a carbapenem-resistant Klebsiella pneumoniae strain, PittNDM01, which coproduces NDM-1 and OXA-232 carbapenemases, was determined in this study. The use of single-molecule, real-time (SMRT) sequencing provided a closed genome in a single sequencing run. K. pneumoniae PittNDM01 has a single chromosome of 5,348,284 bp and four plasmids: pPKPN1 (283,371 bp), pPKPN2 (103,694 bp), pPKPN3 (70,814 bp), and pPKPN4 (6,141 bp). The contents of the chromosome were similar to that of the K. pneumoniae reference genome strain MGH 78578, with the exception of a large inversion spanning 23.3% of the chromosome. In contrast, three of the four plasmids are unique. The plasmid pPKPN1, an IncHI1B-like plasmid, carries the blaNDM-1, armA, and qnrB1 genes, along with tellurium and mercury resistance operons. blaNDM-1 is carried on a unique structure in which Tn125 is further bracketed by IS26 downstream of a class 1 integron. The IncFIA-like plasmid pPKPN3 also carries an array of resistance elements, including blaCTX-M-15 and a mercury resistance operon. The ColE-type plasmid pPKPN4 carrying blaOXA-232 is identical to a plasmid previously reported from France. SMRT sequencing was useful in resolving the complex bacterial genomic structures in the de novo assemblies.
authors: Division of Infectious Diseases, University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania, USA yod4@pitt.edu.
authors: Yohei
authors: Y
authors: Doi
authors: Institute for Genome Sciences, University of Maryland, Baltimore, Maryland, USA.
authors: Tracy H
authors: TH
authors: Hazen
authors: Pacific Biosciences, Menlo Park, California, USA.
authors: Matthew
authors: M
authors: Boitano
authors: Pacific Biosciences, Menlo Park, California, USA.
authors: Yu-Chih
authors: YC
authors: Tsai
authors: Pacific Biosciences, Menlo Park, California, USA.
authors: Tyson A
authors: TA
authors: Clark
authors: Pacific Biosciences, Menlo Park, California, USA.
authors: Jonas
authors: J
authors: Korlach
authors: Institute for Genome Sciences, University of Maryland, Baltimore, Maryland, USA.
authors: David A
authors: DA
authors: Rasko
mesh_terms: D000900:Anti-Bacterial Agents / Q000494:pharmacology; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism*; D016680:Genome, Bacterial / Q000235:genetics; D007711:Klebsiella pneumoniae / Q000187:drug effects / Q000201:enzymology* / Q000235:genetics*; D008826:Microbial Sensitivity Tests; D009876:Operon / Q000235:genetics; D010957:Plasmids / Q000235:genetics; D001618:beta-Lactamases / Q000235:genetics / Q000378:metabolism*
chemical_list: D000900:Anti-Bacterial Agents; D001426:Bacterial Proteins; D001618:beta-Lactamases; C063912:carbapenemase