Rapid development and field evaluation of a portable CRISPR-based assay for Mpox during the 2025 Sierra Leone outbreak

Abstract

Abstract The large 2025 Mpox clade IIb outbreak in Sierra Leone underscores the urgent need for portable, low-cost diagnostics in decentralized settings. While CRISPR-based assays offer high sensitivity and flexibility, their deployment during active outbreaks remains limited. Here we show the rapid development and field evaluation of Mpox SHINE, a CRISPR–Cas13 assay that integrates lyophilized reagents, ambient-temperature lysis, and automated fluorescence detection on the portable DxHub device. The assay achieves analytical sensitivity down to 10 copies/µL. Clinical validation in Sierra Leone, using 56 clinical specimens, confirms complete concordance with qPCR, demonstrating 100% sensitivity and 100% specificity. Crucially, Mpox SHINE also detects the virus directly from unextracted lesion swabs while maintaining 100% sensitivity and specificity. The mean time-to-result is fast, averaging 11.4 minutes for extracted samples and 27.9 minutes for unextracted samples. These findings demonstrate that CRISPR-based diagnostics translate quickly from genomic sequence to clinically validated, deployable tools within a single outbreak window.