RPA-CRISPR-Cas-Mediated\nDual Lateral Flow Assay for\nthe Point-of-Care Testing of HPV16 and HPV18

The incidence of cervical cancer caused by human papillomavirus\n(HPV) infection has increased in recent years. More than half of all\ncervical cancer cases are due to HPV16 and HPV18 infection, so HPV16\nand HPV18 testing is essential to prevent cervical cancer. HPV testing\nis mainly carried out in hospitals, but it is subject to time and\nspecialized medical facilities. On the other hand, home self-testing\nusing simple diagnostics would present an attractive alternative due\nto privacy and flexibility with regard to time and place, provided\nsufficient sensitivity and specificity can be achieved. In this work,\na dual lateral flow assay based on RPA-CRISPR-Cas12a/13a (named RC-LFA)\nfor HPV detection was described. Taking advantage of the cleavage\nspecificity of Cas12a and Cas13a, a CRISPR-Cas12a/Cas13a system was\ndesigned to detect HPV16 and HPV18. The lateral flow strip with two\ntest lines was designed to suit the CRISPR-Cas12a/Cas13 system. RC-LFA\nachieves rapid and simultaneous detection of HPV16 and HPV18 with\nhigh specificity and sensitivity (10 copies/μL) in about 40\nmin from the extraction of nucleic acid to an instrument-free readout.\nRC-LFA is user-friendly and instrument-free, making it a promising\nmethod for HPV self-tests at home.