Abstract: A multi-target nucleic acid detection kit for human papillomavirus (HPV) typing includes a recombinase polymerase amplification (RPA) primer set, an enzyme and a buffer system for Cas12a-crRNA, and a reporter molecule that displays a signal. In the detection method, a multi-channel microfluidic chip is used as a carrier, and RPA primers are designed for corresponding regions of different HPV subtypes to allow isothermal amplification. A crRNA set is designed for amplicons of different subtypes. The crRNA recognizes an HPV target in a sample, and then activates the CRISPR-Cas12a and cuts a reporter group to release a signal, thereby achieving accurate detection on HPV subtypes. The detection method shows a high sensitivity, a low cost, and easy operations, and is expected to be widely used in the screening of HPV infection.