Molecular characterization of an Escherichia coli clinical isolate that produces both metallo-beta-lactamase VIM-2 and extended-spectrum beta-lactamase GES-7: identification of the In8 integron carrying the blaVIM-2 gene.
OBJECTIVE
We have previously reported the isolation of a VIM-2-producing Escherichia coli clinical isolate and the coexistence of blaVIM-2 and blaGES-7 in the same isolate. The aim of this study was to elucidate the genetic environment of these genes.


METHODS
PCR and sequence analysis were used to identify and analyse the blaVIM-containing integron. The location of the blaVIM-2 and blaGES-7 genes was identified by hybridization experiments on I-CeuI-digested genomic DNA after PFGE.


RESULTS
Only the blaVIM-2 gene has been found on a class I integron, designated In8 (1474 bp). Hybridization with the blaVIM-2, the blaGES-7 and the 16S-23S rRNA probes confirmed the chromosomal location of both genes. The blaVIM-2 probe hybridized with a 710 kb fragment whereas the blaGES-7 probe hybridized with a 144 kb fragment of genomic DNA of the E. coli isolate.


CONCLUSIONS
This report provides evidence for the chromosomal location of both blaVIM-2 and blaGES-7 genes carried by an E. coli clinical isolate. In8, containing blaVIM-2, presents an emerging threat of carbapenem resistance among E. coli isolates in Greece.
Galani
Irene
I
4th Department of Internal Medicine, Molecular Biology Section, University General Hospital ATTIKON Rimini 1, 124 62 Chaidari, Greece.
Souli
Maria
M
Koratzanis
Evangelos
E
Chryssouli
Zoi
Z
Giamarellou
Helen
H
D002876:Chromosomes, Bacterial / Q000235:genetics; D004269:DNA, Bacterial / Q000235:genetics; D016521:Electrophoresis, Gel, Pulsed-Field; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics* / Q000302:isolation & purification; D004927:Escherichia coli Infections / Q000382:microbiology; D029968:Escherichia coli Proteins / Q000235:genetics*; D016680:Genome, Bacterial / Q000235:genetics; D006801:Humans; D032023:Integrons / Q000235:genetics*; D015335:Molecular Probes; D008969:Molecular Sequence Data; D009693:Nucleic Acid Hybridization; D016133:Polymerase Chain Reaction; D012336:RNA, Ribosomal, 16S / Q000235:genetics; D012338:RNA, Ribosomal, 23S / Q000235:genetics; D017422:Sequence Analysis, DNA; D001618:beta-Lactamases / Q000096:biosynthesis / Q000235:genetics*
D004269:DNA, Bacterial; D029968:Escherichia coli Proteins; D015335:Molecular Probes; D012336:RNA, Ribosomal, 16S; D012338:RNA, Ribosomal, 23S; C425108:beta-lactamase bla(vim-2); D001618:beta-Lactamases