Molecular characterisation of the recA locus in clinical isolates of verocytotoxigenic E. coli O157:H7

J. E. Moore, M. Watabe, B. C. Millar, P. J. Rooney, A. Loughrey, C. E. Goldsmith, M. A. S. McMahon, D.A McDowell, P. G. Murphy

    Research output: Contribution to journalArticle

    1 Citation (Scopus)

    Abstract

    Molecular epidemiology of verocytoxigenic Escherichia coli O157:H7 is important to help elucidate reservoirs and modes of transmission, particularly between animals and humans. As the recA gene locus is now beginning to gain application in bacterial genotyping schemes, and as it has not been examined previously in E. coli O157 isolates, this study aims to examine potential polymorphic variation as a possible epidemiological marker for the subspecies characterisation of clinically significant verocytotoxigenic E. coli O157:H7. A novel polymerase chain reaction (PCR) assay was designed to target a 638 bp region of the recA gene in E. coli O157 isolates. The PCR amplification of genomic DNA from extracted organisms was able to generate an amplicon of the expected size (approximately 638 bp) for all E. coli O157:H7 examined (n=80), as well as for other non-O157 F. coli and other members of the Enterobacteriaeceae including Citrobacter, Hafnia, Shigella, Enterobacter and Providencia. Subsequent restriction fragment length polymorphism (RFLP) and single-stranded conformational polymorphism (SSCP) analyses of these recA amplicons were able to differentiate E. coli O157 from the organisms examined, but were unable to distinguish between 79 isolates of wild-type E. coli O157, suggesting a highly conserved recA gene structure within the local population of organisms examined.
    LanguageEnglish
    Pages37-41
    JournalBritish Journal of Biomedical Science
    Volume66
    Issue number1
    Publication statusPublished - 2009

    Fingerprint

    Escherichia coli O157
    Hafnia
    Providencia
    Citrobacter
    Genes
    Single-Stranded Conformational Polymorphism
    Polymerase Chain Reaction
    Enterobacter
    Shigella
    Molecular Epidemiology
    Restriction Fragment Length Polymorphisms
    DNA
    Population

    Keywords

    • Escherichia coli O157-H7
    • Genes
    • Polymerase chain reaction
    • Polymorphism
    • restriction fragment length
    • single-stranded conformational

    Cite this

    Moore, J. E., Watabe, M., Millar, B. C., Rooney, P. J., Loughrey, A., Goldsmith, C. E., ... Murphy, P. G. (2009). Molecular characterisation of the recA locus in clinical isolates of verocytotoxigenic E. coli O157:H7. British Journal of Biomedical Science, 66(1), 37-41.
    Moore, J. E. ; Watabe, M. ; Millar, B. C. ; Rooney, P. J. ; Loughrey, A. ; Goldsmith, C. E. ; McMahon, M. A. S. ; McDowell, D.A ; Murphy, P. G. / Molecular characterisation of the recA locus in clinical isolates of verocytotoxigenic E. coli O157:H7. In: British Journal of Biomedical Science. 2009 ; Vol. 66, No. 1. pp. 37-41.
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    abstract = "Molecular epidemiology of verocytoxigenic Escherichia coli O157:H7 is important to help elucidate reservoirs and modes of transmission, particularly between animals and humans. As the recA gene locus is now beginning to gain application in bacterial genotyping schemes, and as it has not been examined previously in E. coli O157 isolates, this study aims to examine potential polymorphic variation as a possible epidemiological marker for the subspecies characterisation of clinically significant verocytotoxigenic E. coli O157:H7. A novel polymerase chain reaction (PCR) assay was designed to target a 638 bp region of the recA gene in E. coli O157 isolates. The PCR amplification of genomic DNA from extracted organisms was able to generate an amplicon of the expected size (approximately 638 bp) for all E. coli O157:H7 examined (n=80), as well as for other non-O157 F. coli and other members of the Enterobacteriaeceae including Citrobacter, Hafnia, Shigella, Enterobacter and Providencia. Subsequent restriction fragment length polymorphism (RFLP) and single-stranded conformational polymorphism (SSCP) analyses of these recA amplicons were able to differentiate E. coli O157 from the organisms examined, but were unable to distinguish between 79 isolates of wild-type E. coli O157, suggesting a highly conserved recA gene structure within the local population of organisms examined.",
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    Moore, JE, Watabe, M, Millar, BC, Rooney, PJ, Loughrey, A, Goldsmith, CE, McMahon, MAS, McDowell, DA & Murphy, PG 2009, 'Molecular characterisation of the recA locus in clinical isolates of verocytotoxigenic E. coli O157:H7', British Journal of Biomedical Science, vol. 66, no. 1, pp. 37-41.

    Molecular characterisation of the recA locus in clinical isolates of verocytotoxigenic E. coli O157:H7. / Moore, J. E.; Watabe, M.; Millar, B. C.; Rooney, P. J.; Loughrey, A.; Goldsmith, C. E.; McMahon, M. A. S.; McDowell, D.A; Murphy, P. G.

    In: British Journal of Biomedical Science, Vol. 66, No. 1, 2009, p. 37-41.

    Research output: Contribution to journalArticle

    TY - JOUR

    T1 - Molecular characterisation of the recA locus in clinical isolates of verocytotoxigenic E. coli O157:H7

    AU - Moore, J. E.

    AU - Watabe, M.

    AU - Millar, B. C.

    AU - Rooney, P. J.

    AU - Loughrey, A.

    AU - Goldsmith, C. E.

    AU - McMahon, M. A. S.

    AU - McDowell, D.A

    AU - Murphy, P. G.

    PY - 2009

    Y1 - 2009

    N2 - Molecular epidemiology of verocytoxigenic Escherichia coli O157:H7 is important to help elucidate reservoirs and modes of transmission, particularly between animals and humans. As the recA gene locus is now beginning to gain application in bacterial genotyping schemes, and as it has not been examined previously in E. coli O157 isolates, this study aims to examine potential polymorphic variation as a possible epidemiological marker for the subspecies characterisation of clinically significant verocytotoxigenic E. coli O157:H7. A novel polymerase chain reaction (PCR) assay was designed to target a 638 bp region of the recA gene in E. coli O157 isolates. The PCR amplification of genomic DNA from extracted organisms was able to generate an amplicon of the expected size (approximately 638 bp) for all E. coli O157:H7 examined (n=80), as well as for other non-O157 F. coli and other members of the Enterobacteriaeceae including Citrobacter, Hafnia, Shigella, Enterobacter and Providencia. Subsequent restriction fragment length polymorphism (RFLP) and single-stranded conformational polymorphism (SSCP) analyses of these recA amplicons were able to differentiate E. coli O157 from the organisms examined, but were unable to distinguish between 79 isolates of wild-type E. coli O157, suggesting a highly conserved recA gene structure within the local population of organisms examined.

    AB - Molecular epidemiology of verocytoxigenic Escherichia coli O157:H7 is important to help elucidate reservoirs and modes of transmission, particularly between animals and humans. As the recA gene locus is now beginning to gain application in bacterial genotyping schemes, and as it has not been examined previously in E. coli O157 isolates, this study aims to examine potential polymorphic variation as a possible epidemiological marker for the subspecies characterisation of clinically significant verocytotoxigenic E. coli O157:H7. A novel polymerase chain reaction (PCR) assay was designed to target a 638 bp region of the recA gene in E. coli O157 isolates. The PCR amplification of genomic DNA from extracted organisms was able to generate an amplicon of the expected size (approximately 638 bp) for all E. coli O157:H7 examined (n=80), as well as for other non-O157 F. coli and other members of the Enterobacteriaeceae including Citrobacter, Hafnia, Shigella, Enterobacter and Providencia. Subsequent restriction fragment length polymorphism (RFLP) and single-stranded conformational polymorphism (SSCP) analyses of these recA amplicons were able to differentiate E. coli O157 from the organisms examined, but were unable to distinguish between 79 isolates of wild-type E. coli O157, suggesting a highly conserved recA gene structure within the local population of organisms examined.

    KW - Escherichia coli O157-H7

    KW - Genes

    KW - Polymerase chain reaction

    KW - Polymorphism

    KW - restriction fragment length

    KW - single-stranded conformational

    M3 - Article

    VL - 66

    SP - 37

    EP - 41

    JO - British Journal of Biomedical Science

    T2 - British Journal of Biomedical Science

    JF - British Journal of Biomedical Science

    SN - 0967-4845

    IS - 1

    ER -

    Moore JE, Watabe M, Millar BC, Rooney PJ, Loughrey A, Goldsmith CE et al. Molecular characterisation of the recA locus in clinical isolates of verocytotoxigenic E. coli O157:H7. British Journal of Biomedical Science. 2009;66(1):37-41.