Publications
Nature biomedical engineeringAug 2022 |
6
(
8
),
944-956
DOI:
10.1038/s41551-022-00917-y

Rapid detection of SARS-CoV-2 RNA in saliva via Cas13

Chandrasekaran, Sita S; Agrawal, Shreeya; Fanton, Alison; Jangid, Aditya R; Charrez, Bérénice; Escajeda, Arturo M; Son, Sungmin; Mcintosh, Roger; Tran, Huyen; Bhuiya, Abdul; de León Derby, María Díaz; Switz, Neil A; Armstrong, Maxim; Harris, Andrew R; Prywes, Noam; Lukarska, Maria; Biering, Scott B; Smock, Dylan C J; Mok, Amanda; Knott, Gavin J; Dang, Qi; Van Dis, Erik; Dugan, Eli; Kim, Shin; Liu, Tina Y; IGI Testing Consortium, ; Moehle, Erica A; Kogut, Katherine; Eskenazi, Brenda; Harris, Eva; Stanley, Sarah A; Lareau, Liana F; Tan, Ming X; Fletcher, Daniel A; Doudna, Jennifer A; Savage, David F; Hsu, Patrick D
Product Used
NGS
Abstract
Rapid nucleic acid testing is central to infectious disease surveillance. Here, we report an assay for rapid COVID-19 testing and its implementation in a prototype microfluidic device. The assay, which we named DISCoVER (for diagnostics with coronavirus enzymatic reporting), involves extraction-free sample lysis via shelf-stable and low-cost reagents, multiplexed isothermal RNA amplification followed by T7 transcription, and Cas13-mediated cleavage of a quenched fluorophore. The device consists of a single-use gravity-driven microfluidic cartridge inserted into a compact instrument for automated running of the assay and readout of fluorescence within 60 min. DISCoVER can detect severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) in saliva with a sensitivity of 40 copies μl-1, and was 94% sensitive and 100% specific when validated (against quantitative PCR) using total RNA extracted from 63 nasal-swab samples (33 SARS-CoV-2-positive, with cycle-threshold values of 13-35). The device correctly identified all tested clinical saliva samples (10 SARS-CoV-2-positive out of 13, with cycle-threshold values of 23-31). Rapid point-of-care nucleic acid testing may broaden the use of molecular diagnostics.
Product Used
NGS

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