dGentamicin doses of 2 and 10 g/ml were bactericidal against 64% and 100%, respectively, of gentamicin-susceptible KPC-2-producing Klebsiella pneumoniae strains. Treatment with the combination of doripenem (8 g/ml) plus colistin (2 g/ml) was inferior to treatment with gentamicin (2 g/ml), doripenem-gentamicin, gentamicin-colistin, and doripenem-gentamicin-colistin against strains with glycine and aspartic acid insertions in OpmK36 porin at amino acid (aa) positions 134 and 135 (n ؍ 9). Doripenem-colistin was comparable to other 2-or 3-drug regimens and superior to single drugs against wild-type/minor ompK36 mutants (n ؍ 5). An algorithm incorporating ompK36 genotypes and susceptibility to gentamicin and doripenem may predict antimicrobial activity against KPC-producing K. pneumoniae.
Carbapenem-resistant Klebsiella pneumoniae strains have emerged as major nosocomial pathogens capable of causing infections that are generally unresponsive to conventional antimicrobial therapy and are associated with high mortality rates (1, 2). Observational studies from our center and others suggest that outcomes are improved with carbapenem-containing combination regimens (3). However, these findings have not been validated in clinical trials, and the optimal combinations have not been defined.Carbapenem resistance is mediated through several mechanisms, including the production of metallo--lactamases and non-metallo-carbapenemases (such as Klebsiella pneumoniae carbapenemase [KPC] and OXA-type carbapenemase), with or without disturbances of outer membrane proteins (OMPs), such as porins. KPC subtype 2 (KPC-2)-producing sequence type 258 (ST258) K. pneumoniae strains predominate in U.S. hospitals and have spread worldwide. At our center and many others, ST258 K. pneumoniae strains carry a mutant ompK35 porin gene, which results in a premature stop codon at amino acid (aa) position 89 (STOP-aa89) (4-6). We previously demonstrated that strains with the mutant ompK35 gene and a wild-type or minor mutant ompK36 gene were highly susceptible to the combination of doripenem and colistin (DORϩCOL) in time-kill assays in vitro. In contrast, DORϩCOL was inactive against strains carrying the mutant ompK35 and major ompK36 mutations, such as an IS5 insertion in the promoter and a 6-bp insertion that encodes glycine and aspartic acid at amino acid positions 134 and 135 (ins aa134-135GD) (7). The ins aa134-135GD mutation is particularly important since the mutated site falls within a transmembrane -strand loop 3 that constitutes the porin channel eyelet. The ins aa134-135GD mutant strains exhibit diminished carbapenem uptake due to porin channel constriction (8).In an earlier time-kill study, we demonstrated that treatments with doripenem and gentamicin (DORϩGENT) were ineffective against GENT-resistant KPC-producing K. pneumoniae (KPC-K. pneumoniae) strains (9). At our center, however, ϳ60% of ST258 KPC-K. pneumoniae strains are GENT susceptible (6). The objectives of this study were to compare the in vitro activities of GENT, DORϩGEN...