Molecular Detection Methods in HPV-Related Cancers

Abstract

Human papillomavirus (HPV) is responsible for most cervical cancers and some head and neck cancers, including oropharyngeal squamous cell carcinoma and sinonasal carcinoma. Cervical cancer is commonly diagnosed by liquid-based cytology, followed by HPV testing using commercially available DNA polymerase chain reaction (PCR), p16 immunohistochemistry (IHC), or DNA/RNA in situ hybridization. HPV in head and neck cancers is commonly diagnosed by p16 IHC or by RT-qPCR of HPV-16 E6 and E7 oncoproteins. Droplet digital PCR has been reported as an ultrasensitive and highly precise method of nucleic acid quantification for biomarker analysis and has been used to detect oncogenic HPV in oropharyngeal and cervical cancers.

Keywords: diagnostic tools; droplet digital polymerase chain reaction; human papillomavirus; immunohistochemistry; in situ hybridization; p16; polymerase chain reaction.

Figure 1

The ddPCR sample is partitioned into 20,000 uniform, nanoliter-sized droplets, and the target and background DNA are distributed randomly into the droplets. (93).

Figure 2

The droplet reader reads spaced-out individual droplet fluorescence in two channels in positive droplets with at least one copy of the target DNA molecule demonstrating increased fluorescence compared to negative droplets. (93).