Triple-mode point-of-care diagnostics for high-risk human papillomavirus in urine

Human papillomavirus (HPV) screening is crucial for early diagnosis and prevention of cervical cancer, yet fast and convenient HPV detection remains challenging, especially in resource-limited areas. Herein, we developed a nucleic acid test named SSMG-LAMP, which combined loop-mediated isothermal amplification (LAMP) with an engineered DNA indicator (SYBR Safe – Malachite Green) for the point-of-care diagnosis of high-risk HPV strains (HPV 16 and 18) in urine. The assay can be completed within 45 min, including DNA extraction, SSMG-LAMP reaction, and signal readout using a simple, portable system. This system enabled the triple-mode detection of DNA targets using colorimetry, fluorometry, and electrochemistry, and can detect as low as 10 copies μL⁻¹ HPV DNA. As a preliminary validation, we used SSMG-LAMP for a blind test of 16 clinical urine samples to detect HPV 16 and 18, and results showed a sensitivity of >80 % and specificity of up to 96.2 %, with a 95 % confidence interval. This triple-mode HPV detection strategy holds potential for point-of-care cervical cancer screening in low-resource settings.