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    The commonly-used DNA probe for diffusely-adherent Escherichia coli cross-reacts with a subset of enteroaggregative E. coli.

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    1471-2180-9-269.pdf (324.1Kb)
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    Publication date
    21/12/2009
    Author
    Snelling, Anna M.
    Macfarlane-Smith, Louissa
    Fletcher, Jonathan N.
    Okeke, Iruka N.
    Keyword
    Diffusely-adherent Escherichia coli (DAEC)
    Enteroaggregative E. coli (EAEC)
    Diarrhoea
    DNA probes
    A HEp-2 adherence assay
    DaaC probe
    PCR-RFLP
    Rights
    © 2009 Snelling et al; licensee BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
    Peer-Reviewed
    Yes
    
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    Abstract
    Background. The roles of diffusely-adherent Escherichia coli (DAEC) and enteroaggregative E. coli (EAEC) in disease are not well understood, in part because of the limitations of diagnostic tests for each of these categories of diarrhoea-causing E. coli. A HEp-2 adherence assay is the Gold Standard for detecting both EAEC and DAEC but DNA probes with limited sensitivity are also employed. Results. We demonstrate that the daaC probe, conventionally used to detect DAEC, cross-reacts with a subset of strains belonging to the EAEC category. The cross hybridization is due to 84% identity, at the nucleotide level, between the daaC locus and the aggregative adherence fimbriae II cluster gene, aafC, present in some EAEC strains. Because aaf-positive EAEC show a better association with diarrhoea than other EAEC, this specific cross-hybridization may have contributed to an over-estimation of the association of daaC with disease in some studies. We have developed a discriminatory PCR-RFLP protocol to delineate EAEC strains detected by the daaC probe in molecular epidemiological studies. Conclusions. A PCR-RFLP protocol described herein can be used to identify aaf-positive EAEC and daaC-positive DAEC and to delineate these two types of diarrhoeagenic E. coli, which both react with the daaC probe. This should help to improve current understanding and future investigations of DAEC and EAEC epidemiology.
    URI
    http://hdl.handle.net/10454/4778
    Version
    published version paper
    Citation
    Snelling, A. M., Macfarlane-Smith, L. R., Fletcher, J. N. and Okeke, I. N. (2009). The commonly-used DNA probe for diffusely-adherent Escherichia coli cross-reacts with a subset of enteroaggregative E. coli. BMC Microbiology, Vol. 9, p. 269 (10p.).
    Link to publisher’s version
    http://www.biomedcentral.com/1471-2180/9/269
    Type
    Article
    Collections
    Life Sciences Publications

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