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Research Article

Association of two resistance mechanisms in a clinical isolate of Enterobacter cloacae with high-level resistance to imipenem.

E H Lee, M H Nicolas, M D Kitzis, G Pialoux, E Collatz, L Gutmann
E H Lee
Laboratoire de Microbiologie Médicale, Université Paris VI, France.
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M H Nicolas
Laboratoire de Microbiologie Médicale, Université Paris VI, France.
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M D Kitzis
Laboratoire de Microbiologie Médicale, Université Paris VI, France.
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G Pialoux
Laboratoire de Microbiologie Médicale, Université Paris VI, France.
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E Collatz
Laboratoire de Microbiologie Médicale, Université Paris VI, France.
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L Gutmann
Laboratoire de Microbiologie Médicale, Université Paris VI, France.
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DOI: 10.1128/AAC.35.6.1093
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ABSTRACT

Carbapenem resistance was studied in a clinical isolate of Enterobacter cloacae, strain 201 (MIC of imipenem and meropenem, 16 micrograms/ml). This strain was analyzed comparatively with the carbapenem-susceptible parent strain 200, an equally susceptible revertant, 201-Rev, and in vitro-selected mutants with different levels of carbapenem resistance. All strains produced similarly high amounts of the same cephalosporinase (pIapp = 8.8). Strain 201 apparently lacked two major outer membrane proteins of ca. 37 and 38 kDa, while 201-Rev produced only the 37-kDa protein. The permeability coefficient, determined with cephaloridine, was reduced up to ninefold in the resistant strains which also showed a substantial reduction in the uptake of [14C]meropenem. The introduction of the plasmid-borne ampD gene (whose product decreases the expression of ampC) resulted in almost complete cessation of cephalosporinase production in all strains and a substantial decrease in the MICs of the carbapenems which remained, however, 8- to 16-fold higher than those determined for the susceptible strains containing the ampD gene. This "residual" resistance was attributed to reduced outer membrane permeability. The contribution of cephalosporinase production was verified in a reverse experiment, in which the introduction of ampC into a low-level cephalosporinase producer resulted in a fourfold increase in the carbapenem MICs. From these results, we infer that reduced outer membrane permeability and high-level cephalosporinase production can operate in conjunction in clinical isolates of E. cloacae to confer imipenem resistance.

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Association of two resistance mechanisms in a clinical isolate of Enterobacter cloacae with high-level resistance to imipenem.
E H Lee, M H Nicolas, M D Kitzis, G Pialoux, E Collatz, L Gutmann
Antimicrobial Agents and Chemotherapy Jun 1991, 35 (6) 1093-1098; DOI: 10.1128/AAC.35.6.1093

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Association of two resistance mechanisms in a clinical isolate of Enterobacter cloacae with high-level resistance to imipenem.
E H Lee, M H Nicolas, M D Kitzis, G Pialoux, E Collatz, L Gutmann
Antimicrobial Agents and Chemotherapy Jun 1991, 35 (6) 1093-1098; DOI: 10.1128/AAC.35.6.1093
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