Sustainable and portable CRISPR-based diagnostics for high-sensitivity Mpox detection

Abstract Mpox has emerged as a critical public health challenge, creating an urgent need for rapid, reliable, and field-deployable diagnostic tools for outbreak settings. Here, we present Kairo-CONAN, a novel CRISPR-Cas3-based point-of-care (POC) diagnostic platform for Mpox, engineered for sustainability and portability. This system leverages a disposable hand warmer (Kairo) as a stable heat source and incorporates freeze-dried reagents for ambient temperature stability, enabling device-free, sensitive detection through lateral flow assay strips. Utilizing CRISPR-Cas3’s unique DNA-targeting and cleavage properties, we optimized probe DNA configurations for high specificity and designed clade-specific target crRNAs. Kairo-CONAN demonstrated rapid, high-sensitivity, and specific detection of Mpox virus (MPXV) DNA across multiple clades, including Clade Ia (Congo), Clade Ib (synthetic DNA), and Clade IIb (Tokyo). By addressing logistical and environmental challenges, Kairo-CONAN offers a sustainable, cost-effective, and field-adapted solution for infectious disease diagnostics, aligning with the 100-day mission framework to enhance global outbreak response efforts.