Virulence typing of Escherichia coli using microarrays

van Ijperen, C.; Kuhnert, Peter; Frey, Joachim; Clewley, J. P. (2002). Virulence typing of Escherichia coli using microarrays. Molecular and cellular probes, 16(5), pp. 371-378. Oxford: Elsevier 10.1006/mcpr.2002.0437

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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.

Item Type:

Journal Article (Original Article)

Division/Institute:

05 Veterinary Medicine > Department of Infectious Diseases and Pathobiology (DIP) > Institute of Veterinary Bacteriology

UniBE Contributor:

Kuhnert, Peter and Frey, Joachim

Subjects:

500 Science
500 Science > 570 Life sciences; biology
600 Technology > 630 Agriculture

ISSN:

0890-8508

Publisher:

Elsevier

Language:

English

Submitter:

Peter Kuhnert-Ryser

Date Deposited:

30 Jan 2014 11:06

Last Modified:

10 Aug 2015 10:35

Publisher DOI:

10.1006/mcpr.2002.0437

PubMed ID:

12477441

Uncontrolled Keywords:

pathogenic Escherichia coli, microarray, virulence typing, probe hybridisation, genetic diversity

BORIS DOI:

10.7892/boris.39125

URI:

https://boris.unibe.ch/id/eprint/39125

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