Laser microdissection for human papillomavirus (HPV) genotyping attribution and methylation pattern analyses of squamous intraepithelial lesions

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.