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Harnessing a novel compact CRISPR-Cas13b for SARS-CoV-2 diagnosticsWang, Qiaochu 04 1900 (has links)
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.
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