Emerging Klebsiella pneumoniae Isolates Coproducing KPC-2 and VIM-1 Carbapenemases

Antimicrobial Agents and Chemotherapy
2009.0

Abstract

Here we report on the emergence of K. pneumoniae isolates coproducing KPC-2 and VIM-1 carbapenemases; to our knowledge, this is the first report of coproduction of VIM and KPC carbapenemases by clinical isolates. Clinical isolates H-1406, A-1797, and T-1780 from hospitals in Crete (Heraklion), Athens, and Thessaly (Trikala), Greece (December 2008 to April 2009) were derived from severely ill patients who had been hospitalized for prolonged time periods (two in intensive care units) and had received multiple courses of antibiotics including carbapenems. Metallo-β-lactamase (ML) production was phenotypically confirmed via EDTA-imipenem synergy, while boronic acid-based tests using imipenem, meropenem, or ertapenem discs appeared negative. PCR and sequencing showed carriage of blaVIM-1 and blaKPC-2 by all three isolates. Their resistance profiles—high-level resistance to penicillins, penicillin-inhibitor combinations, cefotaxime, and ceftazidime, and decreased susceptibility or resistance to cefepime, imipenem, meropenem, and ertapenem—were consistent with VIM-1 and KPC-2 production. Pulsed-field gel electrophoresis of XbaI-digested genomic material showed significant similarity (80%) among the three isolates, distinct from four previously identified KPC-2-producing strains, but isolate T-1780 shared high similarity (95%) with a VIM-1-producing K. pneumoniae strain (A-1760) that has occurred sporadically in Athens hospitals since 2003. Plasmid analysis via the S1 nuclease method revealed that KPC-2 was encoded on similarly sized (~100 kb) transferable plasmids (successfully transferred via conjugation), while VIM-1 was encoded on large (~350 kb) non-transferable plasmids under the tested in vitro conditions. The false-negative results of the boronic acid-based test were attributed to a masking effect of the coproduced VIM-1. Identification of these coproducing isolates in diverse locations within a short time period along with detection difficulties may suggest a thus far unnoticed yet significant spread. Studies are under way to assess the actual prevalence of K. pneumoniae VIM-1 and KPC-2 coproducer isolates and the potential therapeutic impact of the coproduction of VIM-1 and KPC-2 in K. pneumoniae.

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