Performance of a novel, urine-based test for the detection of cervical human papillomavirus infection

Cervical cancer screening using the Pap smear is limited by invasive sampling and low participation among select populations. As human papillomavirus (HPV) is the leading cause of cervical cancer, urine-based testing offers a non-invasive alternative with the potential to increase overall screening participation. We developed and evaluated the performance of the Phase HPV Urine Test, a qualitative real-time PCR assay for the detection of 14 high-risk HPV (HR-HPV) subtypes in first-void urine samples. The assay utilizes a proprietary, room-temperature-stable sample collection system and a novel PHASiFY DNA extraction system optimized for large-volume (40 mL) urine processing. The analytical performance was evaluated to determine accuracy, precision, limit of detection (LOD), cross-reactivity, interference, and sample stability. Analytical performance was also assessed using 128 paired urine and clinician-collected cervical specimens (cervical broom in BD SurePath) tested with the Roche cobas 4800 HPV Test as the reference assay. The assay demonstrated 82.3% initial overall percent agreement with the cobas 4800 HPV test, improving to 93% after adjudication with HPV next-generation sequencing. The assay exhibited high precision (≥97.7%), a low LOD (15-30 viral copies/mL), interference with C. albicans and T. vaginalis, whole blood and peripheral blood mononuclear cells. Urine samples remained stable at room temperature for 10 days, with 95% concordance in results. The Phase HPV Urine Test provides accurate, reproducible, and non-invasive detection of HR-HPV from self-collected urine samples, supporting its potential for expanding access to cervical cancer screening.IMPORTANCEUrine-based testing for high-risk human papillomavirus (HPV) provides a non-invasive, easily accessible alternative that may improve cervical cancer screening participation, especially among under-screened and underserved populations. The Phase HPV Urine Test combines a proprietary urine collection device with a novel large-volume DNA extraction system to enhance the sensitivity and reliability of HPV detection. Our study demonstrates that this approach yields high concordance with standard-of-care testing and maintains sample stability at ambient temperature, enabling flexible, cost-effective transport. By simplifying and standardizing urine collection and by expanding access beyond traditional clinical settings, this technology has the potential to increase uptake of HPV screening programs by overcoming barriers associated with invasive sampling methods and thereby reduce inequities. More importantly, these methods can contribute to earlier detection and prevention of cervical cancer.