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Laser Capture Microdissection pp 167–189Cite as

Laser Microdissection for Human Papillomavirus (HPV) Genotyping Attribution and Methylation Pattern Analyses of Squamous Intraepithelial Lesions

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Part of the book series: Methods in Molecular Biology ((MIMB,volume 1723))

Abstract

Human papillomavirus (HPV) is a nearly ubiquitous infectious organism. It is estimated that 80% of sexually active adults will be exposed to anogenital HPVs in their lifetime, and detection of multiple genotypes in an anogenital sample is common. Detection and genotyping of HPV is usually performed by DNA testing, and less frequently by mRNA testing. HPV genotype testing and characterization of DNA methylation patterns of HPV-related lesions can provide important biological, epidemiological, and potentially relevant clinical information in individuals and populations. The use of laser capture microdissection to isolate cells within a specific lesion allows for very precise molecular characterization and hence causal attribution. This chapter describes detailed protocols for the capture of lesion-specific tissue from formalin-fixed, paraffin-embedded (FFPE) biopsy tissue, and downstream DNA testing for lesion-specific HPV genotype and their methylation patterns.

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Acknowledgments

The authors gratefully thank Dr. Wim Quint, Prof. Magnus von Knebel Doeberitz and Dr. Svetlana Vinokurova for excellent advice and assistance with protocols for laser capture microdissection and bisulfite conversion. We thank also Dr. David Chandler for assistance with pyrosequencing and analysis.

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Authors and Affiliations

  1. Regional HPV Lab Net Reference Laboratory, Department of Microbiology and Infectious Diseases, The Royal Women’s Hospital, Parkville, VIC, Australia

    Monica Molano, Suzanne M. Garland & Alyssa M. Cornall

  2. Murdoch Childrens Research Institute, Parkville, VIC, Australia

    Suzanne M. Garland & Alyssa M. Cornall

  3. Department of Obstetrics and Gynaecology, University of Melbourne, Parkville, VIC, Australia

    Suzanne M. Garland & Alyssa M. Cornall

Authors
  1. Monica Molano
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  2. Suzanne M. Garland
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  3. Alyssa M. Cornall
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Corresponding author

Correspondence to Alyssa M. Cornall .

Editor information

Editors and Affiliations

  1. School of Medicine, Medical Sciences and Nutrition, University of Aberdeen, Aberdeen, United Kingdom

    Graeme I. Murray

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Molano, M., Garland, S.M., Cornall, A.M. (2018). Laser Microdissection for Human Papillomavirus (HPV) Genotyping Attribution and Methylation Pattern Analyses of Squamous Intraepithelial Lesions. In: Murray, G. (eds) Laser Capture Microdissection. Methods in Molecular Biology, vol 1723. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7558-7_9

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  • DOI: https://doi.org/10.1007/978-1-4939-7558-7_9

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-7557-0

  • Online ISBN: 978-1-4939-7558-7

  • eBook Packages: Springer Protocols

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