SlipChip Enables the Integration of CRISPR-Cas12a and RPA for Fast and Stand-Alone HPV Detection

Abstract

Human papillomavirus (HPV) screening is vital for the early detection and prevention of cervical cancer. However, existing methods often face challenges related to speed, simplicity, and multiplexing, especially in resource-limited settings. Here we developed a portable SlipChip-based multiplexed and rapid nucleic acid testing platform, named SMART, designed to simultaneously detect HPV16 and HPV18. SMART allows seamless integration of the RPA and Cas12a assays on the SlipChip and includes a heating membrane to regulate the on-chip assay temperatures. This allows SMART to operate as a stand-alone platform without additional control instruments. The platform also features an All-in-One imaging mode for rapid on-chip data acquisition, enhancing its performance. SMART enables sensitive detection of HPV16 and HPV18 DNA across multiple samples in just 36 min with a detection limit of approximately 6 copies per reaction. Testing of 56 clinical samples at risk of HPV infection validated SMART's performance, showing 97.7% sensitivity and 100% specificity. In summary, SMART offers a stand-alone system capable of rapidly distinguishing between the two most harmful HPV subtypes, showcasing the significant potential for rapid, multiplexed nucleic acid testing in various applications.