Virus detection via programmable Type III-A CRISPR-Cas systems

Among the currently available virus detection assays, those based on the programmable CRISPR-Cas enzymes have the advantage of rapid reporting and high sensitivity without the requirement of thermocyclers. Type III-A CRISPR-Cas system is a multi-component and multipronged immune effector, activated by viral RNA that previously has not been repurposed for disease detection owing in part to the complex enzyme reconstitution process and functionality. Here, we describe the construction and application of a virus detection method, based on an in vivo-reconstituted Type III-A CRISPR-Cas system. This system harnesses both RNA- and transcription-activated dual nucleic acid cleavage activities as well as internal signal amplification that allow virus detection with high sensitivity and at multiple settings. We demonstrate the use of the Type III-A system-based method in detection of SARS-CoV-2 that reached 2000 copies/mu l sensitivity in amplification-free and 60 copies/mu l sensitivity via isothermal amplification within 30 min and diagnosed SARS-CoV-2-infected patients in both settings. The high sensitivity, flexible reaction conditions, and the small molecular-driven amplification make the Type III-A system a potentially unique nucleic acid detection method with broad applications. CRISPR-Cas based virus detection assays can be both rapid and sensitive. Here the authors use a Type III-A CRISPR-Cas system to detect SARS-CoV-2.

Automated summary

CRISPR-Cas-based virus detection assays, particularly those utilizing Type III-A CRISPR-Cas system, offer rapid and sensitive detection of viruses like SARS-CoV-2, making them potentially valuable tools for disease diagnosis.