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Antimicrobial Agents and Chemotherapy, May 2009, p. 1998-2004, Vol. 53, No. 5
0066-4804/09/$08.00+0     doi:10.1128/AAC.01355-08
Copyright © 2009, American Society for Microbiology. All Rights Reserved.

Genetic Organization of Transposase Regions Surrounding blaKPC Carbapenemase Genes on Plasmids from Klebsiella Strains Isolated in a New York City Hospital{triangledown}

Thomas D. Gootz,1* Mary Kay Lescoe,1 Fadia Dib-Hajj,1 Brian A. Dougherty,2 Wen He,2 Phyllis Della-Latta,3 and Richard C. Huard3

Department of Infectious Diseases,1 Molecular Biology, Pfizer Global Research and Development, Groton, Connecticut 06340,2 Clinical Microbiology Service and the Department of Pathology, New York-Presbyterian Hospital, Columbia University Medical Center, New York, New York 100323

Received 9 October 2008/ Returned for modification 12 December 2008/ Accepted 19 February 2009

Carbapenem-resistant Klebsiella strains carrying Klebsiella pneumoniae carbapenemases (KPC) are endemic to New York City and are spreading across the United States and internationally. Recent studies have indicated that the KPC structural gene is located on a 10-kb plasmid-borne element designated Tn4401. Fourteen Klebsiella pneumoniae strains and one Klebsiella oxytoca strain isolated at a New York City hospital in 2005 carrying either blaKPC-2 or blaKPC-3 were examined for isoforms of Tn4401. Ten of the Klebsiella strains contained a 100-bp deletion in Tn4401, corresponding to the Tn4401a isoform. The presence of this deletion adjacent to the upstream promoter region of blaKPC in Tn4401a resulted in a different –35 promoter sequence of TGGAGA than that of CTGATT present in isoform Tn4401b. Complete sequencing of one plasmid carrying blaKPC from each of three nonclonal isolates indicated the presence of genes encoding other types of antibiotic resistance determinants. The 70.6-kb plasmid from K. pneumoniae strain S9 carrying blaKPC-2 revealed two identical copies of Tn4401b inserted in an inverse fashion, but in this case, one of the elements disrupted a group II self-splicing intron. In K. pneumoniae strain S15, the Tn4401a element carrying blaKPC-2 was found on both a large 120-kb plasmid and a smaller 24-kb plasmid. Pulsed-field gel electrophoresis results indicate that the isolates studied represent a heterogeneous group composed of unrelated as well as closely related Klebsiella strains. Our results suggest that endemic KPC-positive Klebsiella strains constitute a generally nonclonal population comprised of various alleles of blaKPC on several distinct plasmid genetic backgrounds. This study increases our understanding of the genetic composition of the evolving and expanding role of KPC-producing, healthcare-associated, gram-negative pathogens.

* Corresponding author. Mailing address: Pfizer Global Research and Development, MS 8118W-211, Building 118W, Groton, CT 06340. Phone: (860) 526-4541. Fax: (860) 715-8162. E-mail: tdgootz{at}yahoo.com

{triangledown} Published ahead of print on 2 March 2009.

Antimicrobial Agents and Chemotherapy, May 2009, p. 1998-2004, Vol. 53, No. 5
0066-4804/09/$08.00+0     doi:10.1128/AAC.01355-08
Copyright © 2009, American Society for Microbiology. All Rights Reserved.



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