Dual-OR Logic-Gated Lateral Flow Strip Assay Based on Colorimetric-Fluorescence Dual Indication for Screening of HPV16/18 in Multiple Scenarios

The incidence of cervical cancer continues to rise in underdeveloped regions due to low human papillomavirus (HPV) vaccination rates and inadequate screening systems. To achieve convenient, rapid, and accurate detection of HPV, we developed a three-wire lateral flow strip assay system based on dual-OR logic gates for rapid and simultaneous detection of HPV subtypes 16 and 18 in a single test. The system combines three-branch-catalytic hairpin assembly (TCHA)-mediated signal amplification with simple OR logic gate-based signal output to improve detection rates while enabling HPV 16/18 subtype identification. The detection limit of the method was calculated to be 10 aM for the selected target sequences. Meanwhile, the method showed excellent specificity with no false-positive output in real-world detection. The sensitivity of the colorimetric test strips exceeded 90%, while the sensitivity of the fluorescence-based test strips surpassed 95% in detecting clinical samples, demonstrating a high degree of concordance with the results obtained from the real-time quantitative polymerase chain reaction (qPCR). This method provides a simple and easy method for the rapid screening of HPV.