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Diagnostic Virology Protocols pp 195–212Cite as

Detection of High-Risk Mucosal Human Papillomavirus DNA in Human Specimens by a Novel and Sensitive Multiplex PCR Method Combined with DNA Microarray

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Part of the book series: Methods in Molecular Biology ((MIMB,volume 665))

Abstract

Epidemiological and functional studies have clearly demonstrated that certain types of human papillomavirus (HPV) from the genus alpha of the HPV phylogenetic tree, referred to as high-risk (HR) types, are the etiological cause of cervical cancer. Several methods for HPV detection and typing have been developed, and their importance in clinical and epidemiological studies has been well demonstrated. However, comparative studies have shown that several assays have different sensitivities for the detection of specific HPV types, particularly in the case of multiple infections.

In this chapter, we describe a novel one-shot method for the detection and typing of 19 mucosal HR HPV types (types 16, 18, 26, 31, 33, 35, 39, 45, 51, 52, 53, 56, 58, 59, 66, 68, 70, 73, and 82). The assay combines the advantages of the multiplex PCR methods, i.e., high sensitivity and the possibility to perform multiple amplifications in a single reaction, with an array primer extension (APEX) assay. The latter method offers the benefits of Sanger dideoxy sequencing with the high-throughput potential of the microarray. Initial studies have revealed that the assay is very sensitive in detecting multiple HPV infections.

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Acknowledgements

We are grateful to all the members of our laboratory for their cooperation and to John Daniel for critical reading of the manuscript. Our research programs are supported by La Ligue Contre le Cancer (Comité du Rhône, Drôme, Savoie), the Association pour la Recherche sur le Cancer, European Union (LSHC-2005-018704), Region Rhône-Alpes and Association for International Cancer Research.

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Authors and Affiliations

  1. Infections and Cancer Biology Group, International Agency for Research on Cancer, Lyon, France

    Tarik Gheit & Massimo Tommasino

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  1. Tarik Gheit
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  2. Massimo Tommasino
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Correspondence to Massimo Tommasino .

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Editors and Affiliations

  1. , Research & Development Division, Health Protection Agency, High Holborn 330, London, WC1V 7BA, United Kingdom

    John R. Stephenson

  2. Northwick Park Hospital, Research and Development, North West London Hospitals NHS Trust, Watford Road, Harrow, HA13UJ, Middlesex, United Kingdom

    Alan Warnes

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Gheit, T., Tommasino, M. (2010). Detection of High-Risk Mucosal Human Papillomavirus DNA in Human Specimens by a Novel and Sensitive Multiplex PCR Method Combined with DNA Microarray. In: Stephenson, J., Warnes, A. (eds) Diagnostic Virology Protocols. Methods in Molecular Biology, vol 665. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-60761-817-1_12

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  • DOI: https://doi.org/10.1007/978-1-60761-817-1_12

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  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-60761-816-4

  • Online ISBN: 978-1-60761-817-1

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