Simplified Cas13-based assays for the fast identification of SARS-CoV-2 and its variants.

TitleSimplified Cas13-based assays for the fast identification of SARS-CoV-2 and its variants.
Publication TypeJournal Article
Year of Publication2022
AuthorsArizti-Sanz, J, Bradley, A'D, Zhang, YB, Boehm, CK, Freije, CA, Grunberg, ME, Kosoko-Thoroddsen, T-SF, Welch, NL, Pillai, PP, Mantena, S, Kim, G, Uwanibe, JN, John, OG, Eromon, PE, Kocher, G, Gross, R, Lee, JS, Hensley, LE, MacInnis, BL, Johnson, J, Springer, M, Happi, CT, Sabeti, PC, Myhrvold, C
JournalNat Biomed Eng
Volume6
Issue8
Pagination932-943
Date Published2022 Aug
ISSN2157-846X
KeywordsCOVID-19, COVID-19 Testing, CRISPR-Associated Proteins, Humans, Nucleic Acids, SARS-CoV-2
Abstract

<p>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.</p>

DOI10.1038/s41551-022-00889-z
Alternate JournalNat Biomed Eng
PubMed ID35637389
PubMed Central IDPMC9398993
Grant ListR01 GM120122 / GM / NIGMS NIH HHS / United States
U01 AI151812 / AI / NIAID NIH HHS / United States
U54 HG007480 / HG / NHGRI NIH HHS / United States
/ HHMI / Howard Hughes Medical Institute / United States