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Virulence typing of Escherichia coli using microarrays.
- Source :
-
Molecular and cellular probes [Mol Cell Probes] 2002 Oct; Vol. 16 (5), pp. 371-8. - Publication Year :
- 2002
-
Abstract
- We describe a microarray based broad-range screening technique for Escherichia coli virulence typing. Gene probes were amplified by PCR from a plasmid bank of characterised E. coli virulence genes and were spotted onto a glass slide to form an array of capture probes. Genomic DNA from E. coli strains which were to be tested for the presence of these virulence gene sequences was labelled with fluorescent cyanine dyes by random amplification and then hybridised against the array of probes. The hybridisation, washing and data analysis conditions were optimised for glass slides, and the applicability of the method for identifying the presence of the virulence genes was determined using reference strains and clinical isolates. It was found to be a sensitive screening method for detecting virulence genes, and a powerful tool for determining the pathotype of E. coli. It will be possible to expand and automate this microarray technique to make it suitable for rapid and reliable diagnostic screening of bacterial isolates.
- Subjects :
- Carbocyanines
DNA Probes genetics
Escherichia coli genetics
Escherichia coli isolation & purification
Escherichia coli Infections diagnosis
Fluorescent Dyes
Glass
Humans
Polymerase Chain Reaction methods
Polymerase Chain Reaction standards
Sensitivity and Specificity
Virulence genetics
Escherichia coli pathogenicity
Oligonucleotide Array Sequence Analysis
Subjects
Details
- Language :
- English
- ISSN :
- 0890-8508
- Volume :
- 16
- Issue :
- 5
- Database :
- MEDLINE
- Journal :
- Molecular and cellular probes
- Publication Type :
- Academic Journal
- Accession number :
- 12477441
- Full Text :
- https://doi.org/10.1006/mcpr.2002.0437