CRISPR-based point-of-care platform for EBV DNA screening and monitoring in nasopharyngeal cancer plasma.

Plasma circulating cell-free Epstein-Barr virus (EBV) DNA is a clinically validated biomarker for early nasopharyngeal carcinoma (NPC) detection and longitudinal disease monitoring. However, current quantitative PCR (qPCR) assays require sophisticated instrumentation and specialized laboratory infrastructure, constraining their accessibility in resource-limited settings. Here, we present two point-of-care testing platforms integrating the RPA-CRISPR/Cas13a system and oil-phase magnetic bead transfer (OMT) method for POCT detection: dμCRISPR, a droplet microfluidic chip for digital quantification, and idCRISPR, an integrated lateral flow device for visual screening. The OMT method achieves efficient and stable EBV DNA extraction from plasma within 8 min. The dμCRISPR platform provides absolute quantitative results within 60 min with an analytical sensitivity of 1 copy/μL. In parallel testing of 24 clinical plasma samples, dμCRISPR were consistent with qPCR. Additionally, the idCRISPR platform enables naked-eye screening within 50 min with a detection limit of 10 copies/μL, showing results consistent with qPCR in the 30 clinical samples evaluated. Together, these platforms form a dual-track strategy for precise quantification and rapid screening, supporting the end-to-end management of NPC across diverse healthcare environments.