Ultrasensitive and visual detection of SARS-CoV-2 using all-in-one dual CRISPR-Cas12a assay

Xiong Ding, Kun Yin, Ziyue Li, Rajesh V. Lalla, Enrique Ballesteros, Maroun M. Sfeir and Changchun Liu ()
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Xiong Ding: University of Connecticut Health Center
Kun Yin: University of Connecticut Health Center
Ziyue Li: University of Connecticut Health Center
Rajesh V. Lalla: University of Connecticut Health Center
Enrique Ballesteros: University of Connecticut Health Center
Maroun M. Sfeir: University of Connecticut Health Center
Changchun Liu: University of Connecticut Health Center

Nature Communications, 2020, vol. 11, issue 1, 1-10

Abstract: Abstract The recent outbreak of novel coronavirus (SARS-CoV-2) causing COVID-19 disease spreads rapidly in the world. Rapid and early detection of SARS-CoV-2 facilitates early intervention and prevents the disease spread. Here, we present an All-In-One Dual CRISPR-Cas12a (AIOD-CRISPR) assay for one-pot, ultrasensitive, and visual SARS-CoV-2 detection. By targeting SARS-CoV-2’s nucleoprotein gene, two CRISPR RNAs without protospacer adjacent motif (PAM) site limitation are introduced to develop the AIOD-CRISPR assay and detect the nucleic acids with a sensitivity of few copies. We validate the assay by using COVID-19 clinical swab samples and obtain consistent results with RT-PCR assay. Furthermore, a low-cost hand warmer (~$0.3) is used as an incubator of the AIOD-CRISPR assay to detect clinical samples within 20 min, enabling an instrument-free, visual SARS-CoV-2 detection at the point of care. Thus, our method has the significant potential to provide a rapid, sensitive, one-pot point-of-care assay for SARS-CoV-2.

Date: 2020
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DOI: 10.1038/s41467-020-18575-6

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