Paper-based CRISPR-Cas diagnostics: A comprehensive review of advances and applications in disease detection

Paper-based analytical devices (PADs) integrated with CRISPR/Cas systems have emerged as innovative tools for point-of-care disease diagnostics, offering simplicity, affordability, and portability. These platforms leverage the high sensitivity and specificity of CRISPR/Cas technology for detecting nucleic acid biomarkers associated with infectious and non-infectious diseases, addressing global healthcare challenges in both resource-limited and wellequipped settings. This review provides a comprehensive overview of paper-based CRISPR/Cas diagnostic platforms, covering detection strategies, device formats (PADs, lateral flow devices, and dipsticks), and recent advancements in isothermal amplification, multiplexed detection, and digital integration. The applications of these platforms in diagnosing bacterial and viral infections, as well as non-infectious diseases such as cancer markers, are discussed in detail, with a focus on innovative approaches to enhance sensitivity, specificity, and usability. Emerging techniques, including amplification-free methods, advanced signal transduction mechanisms, and wearable diagnostic devices, highlight the potential of these platforms to revolutionize healthcare diagnostics. By bridging the gap between laboratory-based testing and real-world applications, paper-based CRISPR/Cas devices represent a promising solution for rapid, accessible, and accurate disease diagnosis, paving the way for future advancements in personalized medicine and global health.