tetano
Editor, Senior Moderator
Nat Biomed Eng
. 2022 May 30.
doi: 10.1038/s41551-022-00889-z. Online ahead of print.
Simplified Cas13-based assays for the fast identification of SARS-CoV-2 and its variants
Jon Arizti-Sanz[SUP] 1 2 [/SUP], A'Doriann Bradley[SUP] 1 [/SUP], Yibin B Zhang[SUP] 1 3 [/SUP], Chloe K Boehm[SUP] 1 4 [/SUP], Catherine A Freije[SUP] 1 [/SUP], Michelle E Grunberg[SUP] 1 4 [/SUP], Tinna-Solveig F Kosoko-Thoroddsen[SUP] 1 [/SUP], Nicole L Welch[SUP] 1 5 [/SUP], Priya P Pillai[SUP] 1 [/SUP], Sreekar Mantena[SUP] 1 3 [/SUP], Gaeun Kim[SUP] 1 4 [/SUP], Jessica N Uwanibe[SUP] 6 7 [/SUP], Oluwagboadurami G John[SUP] 7 [/SUP], Philomena E Eromon[SUP] 6 [/SUP], Gregory Kocher[SUP] 8 [/SUP], Robin Gross[SUP] 8 [/SUP], Justin S Lee[SUP] 9 [/SUP], Lisa E Hensley[SUP] 8 [/SUP], Bronwyn L MacInnis[SUP] 1 10 [/SUP], Jeremy Johnson[SUP] 1 [/SUP], Michael Springer[SUP] 11 12 [/SUP], Christian T Happi[SUP] 6 7 10 [/SUP], Pardis C Sabeti[SUP] #[/SUP][SUP] 1 3 10 13 [/SUP], Cameron Myhrvold[SUP] #[/SUP][SUP] 14 [/SUP]
Affiliations
Abstract
The widespread transmission and evolution of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) call for rapid nucleic acid diagnostics that are easy to use outside of centralized clinical laboratories. Here we report the development and performance benchmarking of Cas13-based nucleic acid assays leveraging lyophilised reagents and fast sample inactivation at ambient temperature. The assays, which we named SHINEv.2 (for 'streamlined highlighting of infections to navigate epidemics, version 2'), simplify the previously reported RNA-extraction-free SHINEv.1 technology by eliminating heating steps and the need for cold storage of the reagents. SHINEv.2 detected SARS-CoV-2 in nasopharyngeal samples with 90.5% sensitivity and 100% specificity (benchmarked against the reverse transcription quantitative polymerase chain reaction) in less than 90 min, using lateral-flow technology and incubation in a heat block at 37 °C. SHINEv.2 also allows for the visual discrimination of the Alpha, Beta, Gamma, Delta and Omicron SARS-CoV-2 variants, and can be run without performance losses by using body heat. Accurate, easy-to-use and equipment-free nucleic acid assays could facilitate wider testing for SARS-CoV-2 and other pathogens in point-of-care and at-home settings.
. 2022 May 30.
doi: 10.1038/s41551-022-00889-z. Online ahead of print.
Simplified Cas13-based assays for the fast identification of SARS-CoV-2 and its variants
Jon Arizti-Sanz[SUP] 1 2 [/SUP], A'Doriann Bradley[SUP] 1 [/SUP], Yibin B Zhang[SUP] 1 3 [/SUP], Chloe K Boehm[SUP] 1 4 [/SUP], Catherine A Freije[SUP] 1 [/SUP], Michelle E Grunberg[SUP] 1 4 [/SUP], Tinna-Solveig F Kosoko-Thoroddsen[SUP] 1 [/SUP], Nicole L Welch[SUP] 1 5 [/SUP], Priya P Pillai[SUP] 1 [/SUP], Sreekar Mantena[SUP] 1 3 [/SUP], Gaeun Kim[SUP] 1 4 [/SUP], Jessica N Uwanibe[SUP] 6 7 [/SUP], Oluwagboadurami G John[SUP] 7 [/SUP], Philomena E Eromon[SUP] 6 [/SUP], Gregory Kocher[SUP] 8 [/SUP], Robin Gross[SUP] 8 [/SUP], Justin S Lee[SUP] 9 [/SUP], Lisa E Hensley[SUP] 8 [/SUP], Bronwyn L MacInnis[SUP] 1 10 [/SUP], Jeremy Johnson[SUP] 1 [/SUP], Michael Springer[SUP] 11 12 [/SUP], Christian T Happi[SUP] 6 7 10 [/SUP], Pardis C Sabeti[SUP] #[/SUP][SUP] 1 3 10 13 [/SUP], Cameron Myhrvold[SUP] #[/SUP][SUP] 14 [/SUP]
Affiliations
- PMID: 35637389
- DOI: 10.1038/s41551-022-00889-z
Abstract
The widespread transmission and evolution of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) call for rapid nucleic acid diagnostics that are easy to use outside of centralized clinical laboratories. Here we report the development and performance benchmarking of Cas13-based nucleic acid assays leveraging lyophilised reagents and fast sample inactivation at ambient temperature. The assays, which we named SHINEv.2 (for 'streamlined highlighting of infections to navigate epidemics, version 2'), simplify the previously reported RNA-extraction-free SHINEv.1 technology by eliminating heating steps and the need for cold storage of the reagents. SHINEv.2 detected SARS-CoV-2 in nasopharyngeal samples with 90.5% sensitivity and 100% specificity (benchmarked against the reverse transcription quantitative polymerase chain reaction) in less than 90 min, using lateral-flow technology and incubation in a heat block at 37 °C. SHINEv.2 also allows for the visual discrimination of the Alpha, Beta, Gamma, Delta and Omicron SARS-CoV-2 variants, and can be run without performance losses by using body heat. Accurate, easy-to-use and equipment-free nucleic acid assays could facilitate wider testing for SARS-CoV-2 and other pathogens in point-of-care and at-home settings.