Automated 5 ' nuclease assay for detection of virulence factors in porcine Escherichia coli

Research output: Contribution to journalJournal article – Annual report year: 2001Researchpeer-review

  • Author: Frydendahl, K.

    Technical University of Denmark

  • Author: Imberechts, H.

  • Author: Lehmann, S.

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This paper describes the development of a 5' nuclease assay for detection of virulence factor genes responsible for colonization factors and toxins in Escherichia coil isolated from pigs. Colonization factors were F4, F5, F6, F18, F41 and the outer membrane protein intimin. Toxins were heat stable (STa, STb, EAST1) and heat labile LT) enterotoxins and the verocytotoxin variant 2e (VT2e). To correctly identify false negative results, an endogenous internal control targeting the E. coil 16S rRNA gene was incorporated in each test tube. The assay was evaluated using a collection of E. coil reference strains which have previously been examined with phenotypical assays or DNA hybridization. Furthermore, the assay was evaluated by testing porcine E. coil field strains, previously characterized. The 5' nuclease assay correctly detected the presence of virulence genes in all reference strains. When testing field strains there was generally excellent agreement with results obtained by laboratories in Belgium and Germany. In conclusion, the 5' nuclease assay developed is a fast and specific tool for detection of E. coli virulence genes in the veterinary diagnostic laboratory.
Original languageEnglish
JournalMolecular and Cellular Probes
Volume15
Issue number3
Pages (from-to)151-160
ISSN0890-8508
DOIs
Publication statusPublished - 2001
CitationsWeb of Science® Times Cited: No match on DOI

    Research areas

  • E. coil, virulence factors, oedema disease, pigs, postweaning diarrhoea
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