The discrimination of high-risk HPV types by in situ hybridization and the polymerase chain reaction

C. S. Herrington, S. M. Anderson, A. K. Graham, James O'D. McGee

    Research output: Contribution to journalArticle

    18 Citations (Scopus)

    Abstract

    The parameter Tm(t) has been defined by non-isotopic in situ hybridization and describes the tissue melting temperature (Tm(t)) of human papillomavirus (HPV) DNA sequences. In this study, multiple in situ hybridization signals for HPV types 16, 31 and 33 in individual archival biopsies hybridized with genomic probes are shown by polymerase chain reactions to be due to cross-hybridization of probe sequences to a single tissue target. Tm(t) is independent of viral type but depends on the homology between probe and target when using nick-translated whole genomic probes. The difference between Tm and Tm(t) is not due to the presence of viral capsid protein. Multiple HPV signals in archival material should not therefore be interpreted as indicative of multiple HPV infection unless adequate stringency conditions have been employed or they are present in morphologically distinct areas of the biopsy.

    Furthermore, extrapolation of calculated DNA homologies to non-isotopic in situ hybridization analysis may not be appropriate. A hybridization signal does not imply probe and target identity: this has implications for HPV typing in clinical material.

    Original languageEnglish
    Pages (from-to)191-198
    Number of pages8
    JournalHistochemical Journal
    Volume25
    Issue number3
    DOIs
    Publication statusPublished - Mar 1993

    Cite this

    Herrington, C. S. ; Anderson, S. M. ; Graham, A. K. ; McGee, James O'D. / The discrimination of high-risk HPV types by in situ hybridization and the polymerase chain reaction. In: Histochemical Journal. 1993 ; Vol. 25, No. 3. pp. 191-198.
    @article{e4a19108ab7040debcff39c49f5c8898,
    title = "The discrimination of high-risk HPV types by in situ hybridization and the polymerase chain reaction",
    abstract = "The parameter Tm(t) has been defined by non-isotopic in situ hybridization and describes the tissue melting temperature (Tm(t)) of human papillomavirus (HPV) DNA sequences. In this study, multiple in situ hybridization signals for HPV types 16, 31 and 33 in individual archival biopsies hybridized with genomic probes are shown by polymerase chain reactions to be due to cross-hybridization of probe sequences to a single tissue target. Tm(t) is independent of viral type but depends on the homology between probe and target when using nick-translated whole genomic probes. The difference between Tm and Tm(t) is not due to the presence of viral capsid protein. Multiple HPV signals in archival material should not therefore be interpreted as indicative of multiple HPV infection unless adequate stringency conditions have been employed or they are present in morphologically distinct areas of the biopsy.Furthermore, extrapolation of calculated DNA homologies to non-isotopic in situ hybridization analysis may not be appropriate. A hybridization signal does not imply probe and target identity: this has implications for HPV typing in clinical material.",
    author = "Herrington, {C. S.} and Anderson, {S. M.} and Graham, {A. K.} and McGee, {James O'D.}",
    year = "1993",
    month = "3",
    doi = "10.1007/BF00163814",
    language = "English",
    volume = "25",
    pages = "191--198",
    journal = "Histochemical Journal",
    issn = "1573-6865",
    publisher = "Springer Verlag",
    number = "3",

    }

    The discrimination of high-risk HPV types by in situ hybridization and the polymerase chain reaction. / Herrington, C. S.; Anderson, S. M.; Graham, A. K.; McGee, James O'D.

    In: Histochemical Journal, Vol. 25, No. 3, 03.1993, p. 191-198.

    Research output: Contribution to journalArticle

    TY - JOUR

    T1 - The discrimination of high-risk HPV types by in situ hybridization and the polymerase chain reaction

    AU - Herrington, C. S.

    AU - Anderson, S. M.

    AU - Graham, A. K.

    AU - McGee, James O'D.

    PY - 1993/3

    Y1 - 1993/3

    N2 - The parameter Tm(t) has been defined by non-isotopic in situ hybridization and describes the tissue melting temperature (Tm(t)) of human papillomavirus (HPV) DNA sequences. In this study, multiple in situ hybridization signals for HPV types 16, 31 and 33 in individual archival biopsies hybridized with genomic probes are shown by polymerase chain reactions to be due to cross-hybridization of probe sequences to a single tissue target. Tm(t) is independent of viral type but depends on the homology between probe and target when using nick-translated whole genomic probes. The difference between Tm and Tm(t) is not due to the presence of viral capsid protein. Multiple HPV signals in archival material should not therefore be interpreted as indicative of multiple HPV infection unless adequate stringency conditions have been employed or they are present in morphologically distinct areas of the biopsy.Furthermore, extrapolation of calculated DNA homologies to non-isotopic in situ hybridization analysis may not be appropriate. A hybridization signal does not imply probe and target identity: this has implications for HPV typing in clinical material.

    AB - The parameter Tm(t) has been defined by non-isotopic in situ hybridization and describes the tissue melting temperature (Tm(t)) of human papillomavirus (HPV) DNA sequences. In this study, multiple in situ hybridization signals for HPV types 16, 31 and 33 in individual archival biopsies hybridized with genomic probes are shown by polymerase chain reactions to be due to cross-hybridization of probe sequences to a single tissue target. Tm(t) is independent of viral type but depends on the homology between probe and target when using nick-translated whole genomic probes. The difference between Tm and Tm(t) is not due to the presence of viral capsid protein. Multiple HPV signals in archival material should not therefore be interpreted as indicative of multiple HPV infection unless adequate stringency conditions have been employed or they are present in morphologically distinct areas of the biopsy.Furthermore, extrapolation of calculated DNA homologies to non-isotopic in situ hybridization analysis may not be appropriate. A hybridization signal does not imply probe and target identity: this has implications for HPV typing in clinical material.

    U2 - 10.1007/BF00163814

    DO - 10.1007/BF00163814

    M3 - Article

    VL - 25

    SP - 191

    EP - 198

    JO - Histochemical Journal

    JF - Histochemical Journal

    SN - 1573-6865

    IS - 3

    ER -