S BEKAL-SI ALI, Biotechnology Research Institute/Université de Montréal sadjia.bekal@nrc.ca
Other Authors
R BROUSSEAU, Biotechnology Research Institute/Université de Montréal L MASSON, Biotechnology Research Institute/Université de Montréal G PRÉFONTAINE, Biotechnology Research Institute/Université de Montréal JM FAIRBROTHER, Université de Montréal J HAREL, Université de Montréal
Title
DNA microarray: a powerful tool for identification of emerging E. coli pathotypes
Abstract Text
Identification of the pathogenicity potential of E. coli strains is mainly based on the detection of virulence genes present in the genome. For this purpose, a DNA microarray was made by spotting DNA amplicons corresponding to 105 virulence genes including adhesin, toxin, invasin, capsul and other virulence-associated genes from human and animal isolates. Attention was given to the choice of the amplicon sequences with regard to similarity and phylogenetic analysis. The usefulness of the microarray was confirmed by a simple hybridization using fluorescently labeled genomic DNA from E. coli strains belonging to defined pathotypes. Assessment of clinical isolates permitted the determination of their pathogenic group and more interestingly allowed the detection of combined pathotypes and the presence of unexpected genes in some strains. The microarray was also shown to be a powerful tool to distinguish among phylogenetic groups using gene probes derived from the attaching and effacing genes (espA, espB and tir). These results showed that the designed virulence microarray is a convenient tool for (i) virulence gene detection (ii) pathotype identification (iii) gene subtyping and (iv) detection of emerging pathotypes. The design of such microarrays could be expanded to other pathogenic bacteria and its applications will be beneficial in the fields of medicine, food quality control and environmental research.
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