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

  • Tarik Gheit
  • Massimo TommasinoEmail author
Protocol
Part of the Methods in Molecular Biology book series (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.

Key words

Cervical cancer High-Risk human papillomaviruses Multiplex PCR Arrayed primer extension Multiple HPV infections 

Notes

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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Copyright information

© Springer Science+Business Media, LLC 2010

Authors and Affiliations

  1. 1.Infections and Cancer Biology GroupInternational Agency for Research on CancerLyonFrance

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