CRISPR‐Based Point‐of‐Care Diagnostics: Cas12, Cas13, Cas14, Microfluidics, and Comparison with PCR

Abstract

Conventional molecular diagnostics, particularly polymerase chain reaction (PCR), remain the gold standard for pathogen detection but are often unsuitable for rapid point‐of‐care (POC) testing because they require specialized equipment and laboratory infrastructure. CRISPR‐Cas systems have emerged as powerful alternatives, offering rapid, accurate, and programmable molecular diagnostics with strong potential for decentralized healthcare. This review summarizes recent advances in CRISPR‐based diagnostics, focusing on Cas12, Cas13, and Cas14, along with key platforms including SHERLOCK, DETECTR, and FELUDA. We discuss their integration with POC technologies such as microfluidic devices, lateral‐flow assays, and smartphone‐based detection systems. Their performance is compared with PCR in terms of sensitivity, specificity, turnaround time, and operational requirements. CRISPR‐based assays demonstrate PCR‐comparable accuracy while enabling isothermal amplification and results within 30–60 minutes. These technologies have supported rapid lateral‐flow testing and received emergency authorization from regulatory agencies, including the U.S. Food and Drug Administration (FDA) and the Indian Council of Medical Research (ICMR), for SARS‐CoV‐2 detection. Despite these advances, challenges such as multiplex detection, reagent stability under ambient conditions, and large‐scale clinical validation remain. Continued improvements in these areas will be essential for translating CRISPR diagnostics into widely accessible, reliable molecular testing at the point of care.