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1.
FEMS Microbiol Lett ; 282(2): 238-40, 2008 May.
Article in English | MEDLINE | ID: mdl-18371063

ABSTRACT

ESAC beta-lactamases have increased catalytic efficiencies toward extended-spectrum cephalosporins and to a lesser extent toward imipenem as compared with the wild-type cephalosporinases. We show here that ESAC expression associated with the loss of both OmpC and OmpF porins conferred in Escherichia coli a high level of resistance to ertapenem and reduced the susceptibility to imipenem. On the contrary, ESAC expressed in the OmpC- or OmpF-deficient E. coli strains or narrow-spectrum cephalosporinase expressed in the OmpC-and OmpF-deficient strain do not confer reduced susceptibility to any of the carbapenems. The production of ESAC beta-lactamase in favorable E. coli background may represent an additional mechanism of resistance to ertapenem.


Subject(s)
Bacterial Proteins/metabolism , Carbapenems/pharmacology , Drug Resistance, Multiple, Bacterial/genetics , Escherichia coli/drug effects , Porins/genetics , beta-Lactamases/metabolism , Anti-Bacterial Agents/metabolism , Anti-Bacterial Agents/pharmacology , Bacterial Proteins/biosynthesis , Carbapenems/metabolism , Cephalosporins/metabolism , Escherichia coli/enzymology , Imipenem/metabolism , Porins/isolation & purification , beta-Lactamases/biosynthesis
2.
J Antimicrob Chemother ; 61(3): 498-503, 2008 Mar.
Article in English | MEDLINE | ID: mdl-18250231

ABSTRACT

OBJECTIVES: To characterize the AmpC-type beta-lactamases produced by Escherichia coli clinical isolates. METHODS: E. coli isolates recovered in a French hospital in 2006 were selected on the basis of a resistance phenotype consistent with increased AmpC production. The presence of genes coding for plasmid-mediated cephalosporinases as well as the existence of mutations in the chromosome-borne ampC genes was studied by PCR and sequencing. Genes for chromosomal cephalosporinases were cloned and the conferred resistance patterns were analysed. The isolates were submitted to phylotyping and genotyping analysis. RESULTS: Thirty-four out of 2800 E. coli isolates were selected. Sixteen isolates, which overexpressed their chromosomal wild-type cephalosporinases due to mutations into their promoter sequence, were susceptible to extended-spectrum cephalosporins (ECLs). Eighteen isolates, mostly of the commensal phylogenetic group A or B1, had reduced susceptibility to ECLs, due to the production of chromosomal extended-spectrum AmpC (ESAC) beta-lactamases, or plasmid-mediated cephalosporinases (CMY-2 and ACC-1), or to combined mechanisms of resistance. Sequence analysis showed that ESAC beta-lactamases had amino acid changes in the R2 binding site, among which was a novel structural change corresponding to the duplication of Ile-283 in the H-9 helix. All the E. coli clinical isolates were non-clonally related except for four CMY-2-producing strains. CONCLUSIONS: This work sheds new light on the spread of ESAC beta-lactamases in E. coli. It showed that this emerging mechanism of resistance could be as frequent as plasmid-mediated cephalosporinases (0.21% and 0.28% of the E. coli isolates, respectively) and that a phenotypic approach is not able to identify these mechanisms of resistance.


Subject(s)
Bacterial Proteins/biosynthesis , Bacterial Proteins/isolation & purification , Escherichia coli/enzymology , Escherichia coli/isolation & purification , Hospitals , beta-Lactamases/biosynthesis , beta-Lactamases/isolation & purification , Bacterial Proteins/genetics , Cross Infection/diagnosis , Cross Infection/enzymology , Cross Infection/microbiology , Escherichia coli/genetics , Escherichia coli Infections/diagnosis , Escherichia coli Infections/enzymology , Escherichia coli Infections/microbiology , France , Humans , Microbial Sensitivity Tests/methods , beta-Lactamases/genetics
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