CRISPR-Cas13 Based Biosensors for Rapid Detection of Viral Pathogens in Point-of-Care Settings

Quick identification of viral pathogens is essential towards enhancing patient outcomes, particularly in point-of-care (POC) settings, where time and accessibility are paramount. Conventional methods of diagnostics are not always sensitive, fast, and portable to use in the field as on-site diagnostics. This paper describes a CRISPR-Cas13-based biosensor that can be used to detect viral pathogens promptly in POC settings and is a fresh implementation of the methodology to improve the diagnostic abilities. A system with the precision and programmability of the CRISPR-Cas13 was paired with a fluorescence-based readout system, allowing direct detection of viral RNA with high specificity. The biosensor design implies the integration of CRISPR-Cas13 that will target particular viral sequences, after which a release of a fluorescence signal will be seen in case of recognition of the target. High sensitivity, fast detection, and ease of use were maximized in the system, which made the system most effective in a decentralized environment, such as a clinic and remote location. The methodology incorporated the creation of a guide RNA specific to the viral genome, conjugation to a small fluorescence reader, and testing in synthetic viral RNA or in clinical samples. The main results show that the CRISPR-Cas13 biosensor is able to identify viral pathogens within less than 30 minutes with a limit of detection (LOD) that matches that of PCR-based approaches. The system was found to be 98 % accurate and could distinguish between related strains of viruses with cross-reactivity. The method is a cost-effective, user-friendly, and fast diagnostic tool that can be used in different settings, giving real-time results on viral infections. The paper emphasizes how CRISPR-Cas13-based biosensors have the potential to transform point-of-care diagnostics by improving health across the globe, especially in under-resourced settings.