Rapid and simultaneous visual typing of high-risk HPV-16/18 with use of integrated lateral flow strip platform

A biosensor for rapid and simultaneous visual identification of high-risk human papillomavirus (HPV) genotypes 16 and 18 in clinical samples based on polymerase chain reaction (PCR) integrated lateral flow strip platform was developed. Using an one-step protocol to extract nucleic acid rapidly and the functionalized primer sets specific to HPV-16 and 18 were designed for the simultaneous amplification. In the presence of target HPV genotypes, the corresponding functionalized primer sets will participate in the PCR process and produce numerous duplex functionalized dsDNA amplicons. With the bridge effect of duplex functionalized dsDNA amplicons between gold nanoparticles-fluorescein isothiocyanate antibody conjugates (AuNP-FITC antibody conjugates) and other two antibodies on corresponding test line (T₁ or T₂), visualized color signals on test lines could be obtained directly visible with a naked eye. Combining the high amplification efficiency of PCR and the visualized sensing of LFS, as low as 700 copies of HPV-16 and 18 DNA were detected simultaneously within 75 min, which can promote application in the resource limited settings. High-risk genotypes of HPV-16 and HPV-18 were easily and simultaneously screened with the amplification-assisted molecular lateral flow strip by on-site observation in the resource-limited settings.