tetano
Editor, Senior Moderator
Nat Biomed Eng
. 2022 Aug 11.
doi: 10.1038/s41551-022-00917-y. Online ahead of print.
Rapid detection of SARS-CoV-2 RNA in saliva via Cas13
Sita S Chandrasekaran[SUP] #[/SUP][SUP] 1 2 3 [/SUP], Shreeya Agrawal[SUP] #[/SUP][SUP] 1 2 [/SUP], Alison Fanton[SUP] #[/SUP][SUP] 1 2 3 [/SUP], Aditya R Jangid[SUP] #[/SUP][SUP] 1 2 [/SUP], Bérénice Charrez[SUP] 3 [/SUP], Arturo M Escajeda[SUP] 4 [/SUP], Sungmin Son[SUP] 1 [/SUP], Roger Mcintosh[SUP] 4 [/SUP], Huyen Tran[SUP] 4 [/SUP], Abdul Bhuiya[SUP] 1 3 [/SUP], María Díaz de León Derby[SUP] 1 3 [/SUP], Neil A Switz[SUP] 5 [/SUP], Maxim Armstrong[SUP] 6 [/SUP], Andrew R Harris[SUP] 1 [/SUP], Noam Prywes[SUP] 2 [/SUP], Maria Lukarska[SUP] 2 7 [/SUP], Scott B Biering[SUP] 8 [/SUP], Dylan C J Smock[SUP] 2 7 [/SUP], Amanda Mok[SUP] 9 [/SUP], Gavin J Knott[SUP] 2 7 10 [/SUP], Qi Dang[SUP] 2 [/SUP], Erik Van Dis[SUP] 7 [/SUP], Eli Dugan[SUP] 2 7 [/SUP], Shin Kim[SUP] 2 7 [/SUP], Tina Y Liu[SUP] 2 7 [/SUP], IGI Testing Consortium; Erica A Moehle[SUP] 2 [/SUP], Katherine Kogut[SUP] 11 [/SUP], Brenda Eskenazi[SUP] 11 [/SUP], Eva Harris[SUP] 8 [/SUP], Sarah A Stanley[SUP] 7 12 [/SUP], Liana F Lareau[SUP] 1 2 [/SUP], Ming X Tan[SUP] 4 [/SUP], Daniel A Fletcher[SUP] 1 [/SUP], Jennifer A Doudna[SUP] 13 14 15 16 17 18 [/SUP], David F Savage[SUP] 19 [/SUP], Patrick D Hsu[SUP] 20 21 22 [/SUP]
Collaborators, Affiliations
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[SUP]-1[/SUP], 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.
. 2022 Aug 11.
doi: 10.1038/s41551-022-00917-y. Online ahead of print.
Rapid detection of SARS-CoV-2 RNA in saliva via Cas13
Sita S Chandrasekaran[SUP] #[/SUP][SUP] 1 2 3 [/SUP], Shreeya Agrawal[SUP] #[/SUP][SUP] 1 2 [/SUP], Alison Fanton[SUP] #[/SUP][SUP] 1 2 3 [/SUP], Aditya R Jangid[SUP] #[/SUP][SUP] 1 2 [/SUP], Bérénice Charrez[SUP] 3 [/SUP], Arturo M Escajeda[SUP] 4 [/SUP], Sungmin Son[SUP] 1 [/SUP], Roger Mcintosh[SUP] 4 [/SUP], Huyen Tran[SUP] 4 [/SUP], Abdul Bhuiya[SUP] 1 3 [/SUP], María Díaz de León Derby[SUP] 1 3 [/SUP], Neil A Switz[SUP] 5 [/SUP], Maxim Armstrong[SUP] 6 [/SUP], Andrew R Harris[SUP] 1 [/SUP], Noam Prywes[SUP] 2 [/SUP], Maria Lukarska[SUP] 2 7 [/SUP], Scott B Biering[SUP] 8 [/SUP], Dylan C J Smock[SUP] 2 7 [/SUP], Amanda Mok[SUP] 9 [/SUP], Gavin J Knott[SUP] 2 7 10 [/SUP], Qi Dang[SUP] 2 [/SUP], Erik Van Dis[SUP] 7 [/SUP], Eli Dugan[SUP] 2 7 [/SUP], Shin Kim[SUP] 2 7 [/SUP], Tina Y Liu[SUP] 2 7 [/SUP], IGI Testing Consortium; Erica A Moehle[SUP] 2 [/SUP], Katherine Kogut[SUP] 11 [/SUP], Brenda Eskenazi[SUP] 11 [/SUP], Eva Harris[SUP] 8 [/SUP], Sarah A Stanley[SUP] 7 12 [/SUP], Liana F Lareau[SUP] 1 2 [/SUP], Ming X Tan[SUP] 4 [/SUP], Daniel A Fletcher[SUP] 1 [/SUP], Jennifer A Doudna[SUP] 13 14 15 16 17 18 [/SUP], David F Savage[SUP] 19 [/SUP], Patrick D Hsu[SUP] 20 21 22 [/SUP]
Collaborators, Affiliations
- PMID: 35953650
- DOI: 10.1038/s41551-022-00917-y
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[SUP]-1[/SUP], 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.