Molecular characterization of AmpC-producing Escherichia coli clinical isolates recovered at two Belgian hospitals.
OBJECTIVE
To characterize the genetic determinants associated with an AmpC phenotype in clinical Escherichia coli isolates.


METHODS
E. coli strains recovered at two Belgian hospitals between 2004 and 2006 were selected on the basis of an AmpC-producing phenotype. Plasmid-mediated cephalosporinases coding genes and the sequence of chromosomal ampC genes were identified by PCR/sequencing. The isolates were submitted to phylotyping and genotyping analysis using rep-PCR (Diversilab) and PFGE. A novel chromosomal ampC gene was cloned.


RESULTS
Eighty-three out of 6850 E. coli isolates were selected. Seventy-two isolates were found to overexpress their chromosomal cephalosporinases while 11 contained plasmid-mediated cephalosporinases. Among chromosomal AmpC overproducers, 12 were extended-spectrum AmpC (ESAC) expressing isolates which all displayed reduced susceptibility to cefepime. Cloning of a new ESAC allele suggested that L293P mutation was responsible of the extension of the hydrolysis spectrum to cefepime. AmpC overproducers, including ESAC producers, predominantly belonged to phylogenetic group A and B1, while plasmid-mediated AmpC-producing isolates preferentially belong to phylogroup B2 and D. According to rep-PCR, the majority of the E. coli isolates belonging to phylogroup A were clonally related which was further confirmed by PFGE for the 11 ESAC expressing isolates.


CONCLUSIONS
Chromosomal AmpC overproduction was the most common resistance mechanism, and the occurrence of ESAC was found to be as frequent as plasmid-mediated cephalosporinases. The detection of a new ESAC allele, of an ESAC producing strain belonging to phylogroup D and the existence of a clonal relationship between ESAC producing strains underline the need for study of the clinical relevance of this mechanism of resistance.
Bogaerts
P
P
Laboratoire de Bactériologie, Cliniques Universitaires UCL de Mont-Godinne, 1 avenue Dr-Gaston-Thérasse, 5530 Yvoir, Belgium. pierre.bogaerts@uclouvain.be
Rodriguez-Villalobos
H
H
Bauraing
C
C
Deplano
A
A
Laurent
C
C
Berhin
C
C
Struelens
M J
MJ
Glupczynski
Y
Y
D001426:Bacterial Proteins / Q000235:genetics*; D001483:Base Sequence; D001530:Belgium / Q000453:epidemiology; D018550:Cephalosporin Resistance / Q000235:genetics*; D002876:Chromosomes, Bacterial / Q000235:genetics; D002999:Clone Cells / Q000187:drug effects / Q000201:enzymology; D003428:Cross Infection / Q000453:epidemiology / Q000382:microbiology*; D016172:DNA Fingerprinting; D004269:DNA, Bacterial / Q000235:genetics; D024901:Drug Resistance, Multiple, Bacterial / Q000235:genetics; D016521:Electrophoresis, Gel, Pulsed-Field; D004926:Escherichia coli / Q000145:classification / Q000187:drug effects / Q000201:enzymology* / Q000302:isolation & purification; D004927:Escherichia coli Infections / Q000453:epidemiology / Q000382:microbiology*; D029968:Escherichia coli Proteins / Q000235:genetics*; D006785:Hospitals, University / Q000706:statistics & numerical data; D006801:Humans; D008969:Molecular Sequence Data; D010802:Phylogeny; D016133:Polymerase Chain Reaction; D011815:R Factors / Q000235:genetics; D017422:Sequence Analysis, DNA; D013379:Substrate Specificity; D001618:beta-Lactamases / Q000235:genetics*
D001426:Bacterial Proteins; D004269:DNA, Bacterial; D029968:Escherichia coli Proteins; C048795:AmpC beta-lactamases; D001618:beta-Lactamases