Hydrogel-based multiplexed high-risk human papillomavirus DNA/RNA detection for cervical cancer screening

Rapid triaging of high-risk human papillomavirus (HPV) infections, especially in resource-limited regions, can reduce cervical cancer deaths by identifying high-risk patients and minimizing pathology bottlenecks and overtreatment. Here, we developed a simple, rapid method for parallel detection of HPV DNA and RNA subtypes associated with a high risk of developing cervical cancer. Specifically, we applied loop-mediated isothermal nucleic acid amplification (LAMP) in a hydrogel-based platform (LAMPGel) for HPV DNA or RNA detection using distinct hydrogel chambers within a single chip. The LAMPGel platform locally restricts single HPV gDNA or mRNA strands with a LAMP cocktail, which is then distributed within a thin hydrogel to locally form single fluorescent spots after a short isothermal incubation of <30 min. Detecting and counting these spots identifies HPV infection, viral load, and, more importantly, RNA expression as a predictive marker for cervical cancer development. LAMPGel is a sensitive, specific, easy-to-use, and rapid method for a point-of-care (POC) platform for potential applications in low- and middle-income countries once the technology is further validated with clinical samples in those settings. Furthermore, it could find broader applications in POC nucleic acid assays.