Harnessing a novel compact CRISPR-Cas13b for SARS-CoV-2 diagnostics

• Main Author: Wang, Qiaochu
• Other Authors: Mahfouz, Magdy M.
• Language: en
• Published: 2021
• Subjects: Diagnostics; SARS-CoV-2; CRISPR-based diagnostics; CRISPR-Cas13; Point-of-care test
• Online Access: Wang, Q. (2021). Harnessing a novel compact CRISPR-Cas13b for SARS-CoV-2 diagnostics. KAUST Research Repository. https://doi.org/10.25781/KAUST-2DDRH; http://hdl.handle.net/10754/669007

The outbreak of infectious diseases across the world results in huge disasters for public health. Rapid and effective diagnostic methods are crucial for disease identification and transmission control. Since first identified in late 2019, the pandemic of COVID-19 caused by the SARS-CoV-2 virus resulted in unprecedented catastrophe globally. To control the further spread of COVID-19, there is an urgent need for rapid, accurate, cost-effective, and efficient diagnostics. Recently, many CRISPR-based diagnostics have been developed by coupling isothermal amplification methods with Cas proteinmediated nucleic acid detection. Compared with conventional methods like RT-qPCR, CRISPR-based assays are more cost-effective and efficient without sacrificing sensitivity and specificity. Here, I developed a Cas13-based assay for SARS-CoV-2 detection with a novel compact Cas13b protein. In this assay, the Cas13 detection is combined with RT-LAMP, achieving the detection of viral RNA as low as 4 copies/μl. By utilizing a simple LED-based visualizer (P51™) instead of a plate reader, the detection result can be visualized directly without using sophisticated instruments. The compact Cas13b-mediated viral detection together with P51™-based visualization enable rapid, sensitive, and portable diagnostics for SARS-CoV-2, showing great potential in application to point-of-care testing.