Development of an integrated microfluidic system for rapid, sensitive HPV detection in plasma samples from head and neck tumor patients: a pilot study
There are approximately 650,000 new cases of head and neck cancer around the world each year. Mucosal human papillomaviruses (HPV) cause cervical cancer and are likely a subset of head and neck squamous cell carcinoma (HNSCC). Researchers from Johns Hopkins University have estimated that 60% of HNSCC patients are HPV positive. HPV detection is vital; positive patients require a different treatment regimen. Previous studies have shown that HPV targets detected in saliva and plasma/serums are potentially useful tools for long-term tumor surveillance of HPV positive oropharyngeal cancer survivors. Currently, issue biopsies are used for HPV detection, but are invasive and inefficient. Circulating cell-free DNA (ccfDNA) are promising, non-invasive candidate biomarkers for early detection, prognosis or treatment selection. Their analysis provides access to the humoral state without the need for tissue biopsy. When cancer cells rupture, they release genome fragments that float freely through the bloodstream. By developing and refining techniques for measuring and sequencing ccfDNA in the bloodstream, scientists can run vials of blood into “liquid biopsies”. In this pilot study, we evaluated five HNSCC patient plasma samples purchased from Conversant Bio (Huntsville, AL) for HPV infection. Using MagJET’s DNA and RNA purification kit (Thermo Scientific), ccfDNA was first isolated from 200μL oropharyngeal HNSCC plasma samples. It was then quantified, qualified, and amplified by real-time isothermal PCR. Four of five samples were HPV 16 positive. Then, indirect anti-E7 ELISA was utilized to confirm the detection of HPV 16. An integrated microfluidic system for rapid and automated HPV detection was designed and initial tests were performed. The work from this study will aid in the development of a rapid, efficient and portable virus detection systems for an array of different cancers and illnesses.