tetano
Editor, Senior Moderator
Nat Commun
. 2020 Sep 18;11(1):4711.
doi: 10.1038/s41467-020-18575-6.
Ultrasensitive and visual detection of SARS-CoV-2 using all-in-one dual CRISPR-Cas12a assay
Xiong Ding[SUP] 1 [/SUP], Kun Yin[SUP] 1 [/SUP], Ziyue Li[SUP] 1 [/SUP], Rajesh V Lalla[SUP] 2 [/SUP], Enrique Ballesteros[SUP] 3 [/SUP], Maroun M Sfeir[SUP] 3 [/SUP], Changchun Liu[SUP] 4 [/SUP]
Affiliations
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.
. 2020 Sep 18;11(1):4711.
doi: 10.1038/s41467-020-18575-6.
Ultrasensitive and visual detection of SARS-CoV-2 using all-in-one dual CRISPR-Cas12a assay
Xiong Ding[SUP] 1 [/SUP], Kun Yin[SUP] 1 [/SUP], Ziyue Li[SUP] 1 [/SUP], Rajesh V Lalla[SUP] 2 [/SUP], Enrique Ballesteros[SUP] 3 [/SUP], Maroun M Sfeir[SUP] 3 [/SUP], Changchun Liu[SUP] 4 [/SUP]
Affiliations
- PMID: 32948757
- DOI: 10.1038/s41467-020-18575-6
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.