Rapid detection of HPV16/18 based on a CRISPR-Cas13a/Cas12a dual-channel system

cervical cancer is the leading cause of cancer deaths in women in the developing world, with high-risk HPV16 and HPV18 accounting for approximately 70% of all cervical cancers. Early detection of HPV, especially high-risk HPV types, is essential to prevent disease progression.in this study, we established a highly sensitive and specific nucleic acid assay based on a CRISPR-Cas13a/Cas12a dual-channel system combined with multiplex RAA for rapid detection and typing of HPV16/18, which provides a new idea for cervical cancer screening. To meet the application of field testing, we designed a portable fluorescence imaging assay that can judge the test results directly with the naked eye or through cell phone imaging.the lower limit of detection for both HPV16 and HPV18 based on the CRISPR-Cas12a/Cas13a dual-channel assay was 100 copies per μL. The dual-channel assay was validated with 55 clinical samples, showing 97.06% sensitivity, 100% specificity, 100% positive predictive value, and 96.55% negative predictive value. The results of the portable fluorescence imaging assay were fully comparable to those of the real-time fluorescent RAA-based CRISPR-Cas12a/Cas13a dual-channel assay.this developed portable dual gene assay platform may provide new technical support for cervical cancer screening in resource-limited settings.