Quantitative real-time PCR detection of Propionibacterium acnes in archived prostate tissue

E. Barnard, S. Patrick, D. Fairley, M. Catherwood, L. Martin, B. Shannon, A. McDowell

    Research output: Chapter in Book/Report/Conference proceedingConference contribution

    Abstract

    The Gram-positive skin commensal Propionibacterium acnes is an opportunistic pathogen associated with, for example, prosthetic joint failure, endocarditis, and ocular infections. P. acnes is frequently present in prostate tissue where it is significantly associated with acute and chronic inflammatory changes. These could potentially stimulate carcinogenesis. Current methods for P. acnes detection are sub-optimal due to slow growth of the organism in culture coupled with the lack of sensitivity of end-point PCRs for low level pathogen detection in clinical samples. We have developed a novel quantitative TaqMan® PCR assay to detect and quantify P. acnes specific 16S rRNA sequences. This assay is sensitive to ten genome equivalents when applied to pure cultures, and exhibits no cross-reactivity when tested against a specificity panel of over 50 species including common contaminants and other members of the Propionibacteriaceae. We have applied this assay to DNA extracted from archived prostate tissue specimens. Normalization with a human endogenous retrovirus-3 (ERV-3) assay has enabled comparison of cancerous prostate, non-cancerous prostate and ‘spiked’ prostate tissue controls. For comparison our assay has also been applied to other clinical samples including failed prosthetic joints, skin and other tissue controls. These data will be discussed in relation to clinical information.
    LanguageEnglish
    Title of host publicationUnknown Host Publication
    Pages74-74
    Number of pages1
    Publication statusPublished - Sep 2011
    EventSociety for General Microbiology Autumn Conference; (YO13) Anaerobe 2011: Anaerobes of the human gastrointestinal microbiota & disease - University of York, UK
    Duration: 1 Sep 2011 → …

    Conference

    ConferenceSociety for General Microbiology Autumn Conference; (YO13) Anaerobe 2011: Anaerobes of the human gastrointestinal microbiota & disease
    Period1/09/11 → …

    Fingerprint

    Propionibacterium acnes
    Real-Time Polymerase Chain Reaction
    Prostate
    Propionibacteriaceae
    Joints
    Endogenous Retroviruses
    Eye Infections
    Polymerase Chain Reaction
    Skin
    Endocarditis
    Carcinogenesis
    Genome
    DNA
    Growth

    Cite this

    Barnard, E., Patrick, S., Fairley, D., Catherwood, M., Martin, L., Shannon, B., & McDowell, A. (2011). Quantitative real-time PCR detection of Propionibacterium acnes in archived prostate tissue. In Unknown Host Publication (pp. 74-74)
    Barnard, E. ; Patrick, S. ; Fairley, D. ; Catherwood, M. ; Martin, L. ; Shannon, B. ; McDowell, A. / Quantitative real-time PCR detection of Propionibacterium acnes in archived prostate tissue. Unknown Host Publication. 2011. pp. 74-74
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    title = "Quantitative real-time PCR detection of Propionibacterium acnes in archived prostate tissue",
    abstract = "The Gram-positive skin commensal Propionibacterium acnes is an opportunistic pathogen associated with, for example, prosthetic joint failure, endocarditis, and ocular infections. P. acnes is frequently present in prostate tissue where it is significantly associated with acute and chronic inflammatory changes. These could potentially stimulate carcinogenesis. Current methods for P. acnes detection are sub-optimal due to slow growth of the organism in culture coupled with the lack of sensitivity of end-point PCRs for low level pathogen detection in clinical samples. We have developed a novel quantitative TaqMan{\circledR} PCR assay to detect and quantify P. acnes specific 16S rRNA sequences. This assay is sensitive to ten genome equivalents when applied to pure cultures, and exhibits no cross-reactivity when tested against a specificity panel of over 50 species including common contaminants and other members of the Propionibacteriaceae. We have applied this assay to DNA extracted from archived prostate tissue specimens. Normalization with a human endogenous retrovirus-3 (ERV-3) assay has enabled comparison of cancerous prostate, non-cancerous prostate and ‘spiked’ prostate tissue controls. For comparison our assay has also been applied to other clinical samples including failed prosthetic joints, skin and other tissue controls. These data will be discussed in relation to clinical information.",
    author = "E. Barnard and S. Patrick and D. Fairley and M. Catherwood and L. Martin and B. Shannon and A. McDowell",
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    Barnard, E, Patrick, S, Fairley, D, Catherwood, M, Martin, L, Shannon, B & McDowell, A 2011, Quantitative real-time PCR detection of Propionibacterium acnes in archived prostate tissue. in Unknown Host Publication. pp. 74-74, Society for General Microbiology Autumn Conference; (YO13) Anaerobe 2011: Anaerobes of the human gastrointestinal microbiota & disease, 1/09/11.

    Quantitative real-time PCR detection of Propionibacterium acnes in archived prostate tissue. / Barnard, E.; Patrick, S.; Fairley, D.; Catherwood, M.; Martin, L.; Shannon, B.; McDowell, A.

    Unknown Host Publication. 2011. p. 74-74.

    Research output: Chapter in Book/Report/Conference proceedingConference contribution

    TY - GEN

    T1 - Quantitative real-time PCR detection of Propionibacterium acnes in archived prostate tissue

    AU - Barnard, E.

    AU - Patrick, S.

    AU - Fairley, D.

    AU - Catherwood, M.

    AU - Martin, L.

    AU - Shannon, B.

    AU - McDowell, A.

    PY - 2011/9

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    N2 - The Gram-positive skin commensal Propionibacterium acnes is an opportunistic pathogen associated with, for example, prosthetic joint failure, endocarditis, and ocular infections. P. acnes is frequently present in prostate tissue where it is significantly associated with acute and chronic inflammatory changes. These could potentially stimulate carcinogenesis. Current methods for P. acnes detection are sub-optimal due to slow growth of the organism in culture coupled with the lack of sensitivity of end-point PCRs for low level pathogen detection in clinical samples. We have developed a novel quantitative TaqMan® PCR assay to detect and quantify P. acnes specific 16S rRNA sequences. This assay is sensitive to ten genome equivalents when applied to pure cultures, and exhibits no cross-reactivity when tested against a specificity panel of over 50 species including common contaminants and other members of the Propionibacteriaceae. We have applied this assay to DNA extracted from archived prostate tissue specimens. Normalization with a human endogenous retrovirus-3 (ERV-3) assay has enabled comparison of cancerous prostate, non-cancerous prostate and ‘spiked’ prostate tissue controls. For comparison our assay has also been applied to other clinical samples including failed prosthetic joints, skin and other tissue controls. These data will be discussed in relation to clinical information.

    AB - The Gram-positive skin commensal Propionibacterium acnes is an opportunistic pathogen associated with, for example, prosthetic joint failure, endocarditis, and ocular infections. P. acnes is frequently present in prostate tissue where it is significantly associated with acute and chronic inflammatory changes. These could potentially stimulate carcinogenesis. Current methods for P. acnes detection are sub-optimal due to slow growth of the organism in culture coupled with the lack of sensitivity of end-point PCRs for low level pathogen detection in clinical samples. We have developed a novel quantitative TaqMan® PCR assay to detect and quantify P. acnes specific 16S rRNA sequences. This assay is sensitive to ten genome equivalents when applied to pure cultures, and exhibits no cross-reactivity when tested against a specificity panel of over 50 species including common contaminants and other members of the Propionibacteriaceae. We have applied this assay to DNA extracted from archived prostate tissue specimens. Normalization with a human endogenous retrovirus-3 (ERV-3) assay has enabled comparison of cancerous prostate, non-cancerous prostate and ‘spiked’ prostate tissue controls. For comparison our assay has also been applied to other clinical samples including failed prosthetic joints, skin and other tissue controls. These data will be discussed in relation to clinical information.

    M3 - Conference contribution

    SP - 74

    EP - 74

    BT - Unknown Host Publication

    ER -

    Barnard E, Patrick S, Fairley D, Catherwood M, Martin L, Shannon B et al. Quantitative real-time PCR detection of Propionibacterium acnes in archived prostate tissue. In Unknown Host Publication. 2011. p. 74-74