Development of an integrated microfluidic system for early HPV detection in head and neck carcinomas
The incidence of head and neck cancer globally is rising steadily at 690,000 new cases annually, ranking at the sixth most common malignancy in the world. Research illustrates approximately 26% of all HNSCC are associated with human papillomaviruses. Studies have indicated that early detection of HPV is clinically significant for prognosis and treatment selection. Liquid biopsy presents many advantages over tissue biopsy; it is minimally invasive, repeatable, and cost-effective. In this study, plasma, saliva, and buccal swab samples from eight HonorHealth Scottsdale HNSCC patients were analyzed for HPV status. The cell-free circulating DNA (ccfDNA) was extracted from the samples using a MagJET DNA and RNA Kit (Thermo Scientific), followed by quantification. A real-time isothermal amplification was performed with HPV-specific primer sets (Atila BioSystems). HPV-16 was detected in all sample types, with 50%, 75%, and 87.5% positivity in plasma, saliva and buccal swab samples, respectively. HPV positive results from HNSCC patients were further confirmed with ELISA for the anti-E7 antibody and all eight plasma samples were positive in this assay. Integration of HPV sample prep and detection in a microfluidic cartridge was attempted. A high yield on-cartridge DNA extraction was achieved; however, the purity needs to be improved. Future work includes optimizing the microfluidic cartridge design and integrating on-chip isothermal amplification with real-time detection.