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.
This publication was prepared for the Directorate-General for Energy, the Directorate-General for Climate
Action and the Directorate-General for Mobility and Transport by the E3M-Lab of the Institute of Communication and Computer Systems at the National Technical University of Athens (ICCS-NTUA), Greece, in
cooperation with the International Institute for Applied Systems Analysis (IIASA) and EuroCARE and represents those organisations’ views on energy, transport and GHG emissions facts, figures and projections.
These views should not be considered as a statement of the Commission’s or the Directorate-General’s