Abstract
The genomes of the human papillomaviruses HPV-16 and HPV-18 undergo increased CpG methylation during the progression of cervical neoplasia, possibly in response to increased recombination between viral and cellular DNA in high-grade lesions. This behavior makes HPV DNA methylation a useful biomarker of carcinogenic progression of HPV infections. The first step in detecting DNA methylation involves modification by bisulfite, which converts cytosine residues into uracil, but leaves 5-methylcytosine residues unaffected. A combination of this reaction with PCR and DNA sequencing permits to evaluate the methylation status of the sample DNA. This chapter describes the basic protocol to measure HPV-16 and HPV-18 CpG methylation by direct sequencing of the PCR products and discusses the value of modified strategies including DNA cloning, amplification with methylation-specific primers, and real-time PCR with TaqMan probes.
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Kalantari, M., Bernard, HU. (2015). Assessment of the HPV DNA Methylation Status in Cervical Lesions. In: Keppler, D., Lin, A. (eds) Cervical Cancer. Methods in Molecular Biology, vol 1249. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-2013-6_20
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DOI: https://doi.org/10.1007/978-1-4939-2013-6_20
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