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Archives of Gynecology and Obstetrics

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01.10.2011 | Gynecologic Oncology

Evaluation of two commercialised in situ hybridisation assays for detecting HPV-DNA in formalin-fixed, paraffin-embedded tissue

verfasst von: Maresa Montag, Thomas J. F. Blankenstein, Naim Shabani, Ansgar Brüning, Ioannis Mylonas

Erschienen in: Archives of Gynecology and Obstetrics | Ausgabe 4/2011

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Abstract

Introduction

The role of human papilloma virus (HPV) in the pathogenesis of anogenital dysplasia is now conclusive. However, HPV detection in formalin-fixed and paraffin-embedded tissues remains controversial. Therefore, the aim of this study was to evaluate morphological changes directly in tissue specimens using a HPV-DNA detection system involving HPV in situ hybridisation.

Materials and methods

Samples from patients with cervical carcinoma were analysed using the GenPoint HPV DNA Probe Cocktail (Dako, Glostrup, Denmark) and the ZytoFast HPV Screening CISH-Kit (Zytomed, Berlin, Germany). Three cervical carcinoma cell lines with a well-defined HPV copy number per cell (SiHa, HeLa, and CaSki) served as positive controls for sensitivity testing, while two HPV-negative cell lines (AC-1M32, MCF-7) and brain tissue samples served as negative controls. Moreover, to assess the validity of the in situ hybridisation, the expression of HPV-16 DNA in cell lines was demonstrated by HPV-16 E6-specific PCR.

Results

Both HPV-screening assays revealed strong signals of episomal and integrated HPV-DNA at a HPV copy number of more than 50 copies/cell. All cervical carcinoma samples were positive in the Dako assay, which identifies 13 high-risk HPV genotypes, whereas HPV-DNA could be detected in 9/10 cervical carcinoma samples using the Zytofast assay, identifying HPV 16, 18, 31, 33, and 35.

Conclusion

HPV in situ hybridisation is a convenient and powerful tool for detecting HPV-DNA in formalin-fixed and paraffin-embedded tissue samples. Therefore, this technique is suitable for analysis of a potential HPV infection using archival pathological slides.

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Titel: Evaluation of two commercialised in situ hybridisation assays for detecting HPV-DNA in formalin-fixed, paraffin-embedded tissue
verfasst von: Maresa Montag
Thomas J. F. Blankenstein
Naim Shabani
Ansgar Brüning
Ioannis Mylonas
Publikationsdatum: 01.10.2011
Verlag: Springer-Verlag
Erschienen in: Archives of Gynecology and Obstetrics / Ausgabe 4/2011
Print ISSN: 0932-0067
Elektronische ISSN: 1432-0711
DOI: https://doi.org/10.1007/s00404-010-1771-z

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Springer Medizin

Evaluation of two commercialised in situ hybridisation assays for detecting HPV-DNA in formalin-fixed, paraffin-embedded tissue

Abstract

Introduction

Materials and methods

Results

Conclusion

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Springer Medizin

Abstract

Introduction

Materials and methods

Results

Conclusion

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