Equipment-free detection of SARS-CoV-2 and Variants of Concern using Cas13.

Autor: Arizti-Sanz J; Broad Institute of Massachusetts Institute of Technology (MIT) and Harvard, Cambridge, MA 02142, USA.; Harvard-MIT Program in Health Sciences and Technology, 77 Massachusetts Avenue, Cambridge, MA 02139, USA., Bradley A; Broad Institute of Massachusetts Institute of Technology (MIT) and Harvard, Cambridge, MA 02142, USA., Zhang YB; Broad Institute of Massachusetts Institute of Technology (MIT) and Harvard, Cambridge, MA 02142, USA.; Department of Organismic and Evolutionary Biology, Harvard University, 26 Oxford Street, Cambridge, MA 02138, USA., Boehm CK; Broad Institute of Massachusetts Institute of Technology (MIT) and Harvard, Cambridge, MA 02142, USA.; Department of Molecular Biology, Princeton University, Princeton, NJ 08544, USA., Freije CA; Broad Institute of Massachusetts Institute of Technology (MIT) and Harvard, Cambridge, MA 02142, USA., Grunberg ME; Broad Institute of Massachusetts Institute of Technology (MIT) and Harvard, Cambridge, MA 02142, USA.; Department of Molecular Biology, Princeton University, Princeton, NJ 08544, USA., Kosoko-Thoroddsen TF; Broad Institute of Massachusetts Institute of Technology (MIT) and Harvard, Cambridge, MA 02142, USA., Welch NL; Broad Institute of Massachusetts Institute of Technology (MIT) and Harvard, Cambridge, MA 02142, USA.; Program in Virology, Harvard Medical School, Boston, MA 02115, USA., Pillai PP; Broad Institute of Massachusetts Institute of Technology (MIT) and Harvard, Cambridge, MA 02142, USA., Mantena S; Broad Institute of Massachusetts Institute of Technology (MIT) and Harvard, Cambridge, MA 02142, USA., Kim G; Broad Institute of Massachusetts Institute of Technology (MIT) and Harvard, Cambridge, MA 02142, USA.; Department of Molecular Biology, Princeton University, Princeton, NJ 08544, USA., Uwanibe JN; African Centre of Excellence for Genomics of Infectious Diseases (ACEGID), Redeemer's University, Ede, Osun State, Nigeria.; Department of Biological Sciences, College of Natural Sciences, Redeemer's University, Ede, Osun State, Nigeria., John OG; Department of Biological Sciences, College of Natural Sciences, Redeemer's University, Ede, Osun State, Nigeria., Eromon PE; African Centre of Excellence for Genomics of Infectious Diseases (ACEGID), Redeemer's University, Ede, Osun State, Nigeria., Kocher G; Integrated Research Facility, Division of Clinical Research, National Institute of Allergy and infectious diseases, National Institute of Health, Frederick, MD 21702, USA., Gross R; Integrated Research Facility, Division of Clinical Research, National Institute of Allergy and infectious diseases, National Institute of Health, Frederick, MD 21702, USA., Lee JS; Biotechnology Cores Facility Branch,Division of Scientific Resources, National Center for Emerging and Infectious Diseases, Centers for Disease Control and Prevention, Atlanta, Georgia, USA., Hensley LE; Integrated Research Facility, Division of Clinical Research, National Institute of Allergy and infectious diseases, National Institute of Health, Frederick, MD 21702, USA., Happi CT; African Centre of Excellence for Genomics of Infectious Diseases (ACEGID), Redeemer's University, Ede, Osun State, Nigeria.; Department of Biological Sciences, College of Natural Sciences, Redeemer's University, Ede, Osun State, Nigeria.; Harvard T.H. Chan School of Public Health, 677 Huntington Avenue, Boston, MA 02115, USA., Johnson J; Broad Institute of Massachusetts Institute of Technology (MIT) and Harvard, Cambridge, MA 02142, USA., Sabeti PC; Broad Institute of Massachusetts Institute of Technology (MIT) and Harvard, Cambridge, MA 02142, USA.; Department of Organismic and Evolutionary Biology, Harvard University, 26 Oxford Street, Cambridge, MA 02138, USA.; Harvard T.H. Chan School of Public Health, 677 Huntington Avenue, Boston, MA 02115, USA.; Howard Hughes Medical Institute, Chevy Chase, MD 20815, USA.; These authors jointly supervised this work: Pardis C. Sabeti, Cameron Myhrvold., Myhrvold C; Department of Molecular Biology, Princeton University, Princeton, NJ 08544, USA.; These authors jointly supervised this work: Pardis C. Sabeti, Cameron Myhrvold.
Jazyk: angličtina
Zdroj: MedRxiv : the preprint server for health sciences [medRxiv] 2021 Nov 02. Date of Electronic Publication: 2021 Nov 02.
DOI: 10.1101/2021.11.01.21265764
Abstrakt: The COVID-19 pandemic, and the recent rise and widespread transmission of SARS-CoV-2 Variants of Concern (VOCs), have demonstrated the need for ubiquitous nucleic acid testing outside of centralized clinical laboratories. Here, we develop SHINEv2, a Cas13-based nucleic acid diagnostic that combines quick and ambient temperature sample processing and lyophilized reagents to greatly simplify the test procedure and assay distribution. We benchmarked a SHINEv2 assay for SARS-CoV-2 detection against state-of-the-art antigen-capture tests using 96 patient samples, demonstrating 50-fold greater sensitivity and 100% specificity. We designed SHINEv2 assays for discriminating the Alpha, Beta, Gamma and Delta VOCs, which can be read out visually using lateral flow technology. We further demonstrate that our assays can be performed without any equipment in less than 90 minutes. SHINEv2 represents an important advance towards rapid nucleic acid tests that can be performed in any location.
Databáze: MEDLINE