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A multiplexed Cas13-based assay with point-of-care attributes for simultaneous COVID-19 diagnosis and variant surveillance
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elife-MS-Fig-SI-V7.docx | Accepted version | 12.17 MB | Microsoft Word | View/Open |
Title: | A multiplexed Cas13-based assay with point-of-care attributes for simultaneous COVID-19 diagnosis and variant surveillance |
Authors: | Patchsung, M Homchan, A Aphicho, K Suraritdechachai, S Wanitchanon, T Pattama, A Sappakhaw, K Meesawat, P Wongsatit, T Athipanyasilp, A Jantarug, K Athipanyasilp, N Buahom, J Visanpattanasin, S Niljianskul, N Chaiyen, P Tinikul, R Wichukchinda, N Mahasirimongkol, S Sirijatuphat, R Angkasekwinai, N Crone, MA Freemont, PS Joung, J Ladha, A Abudayyeh, O Gootenberg, J Zhang, F Chewapreecha, C Chanarat, S Horthongkham, N Pakotiprapha, D Uttamapinant, C |
Item Type: | Journal Article |
Abstract: | Point-of-care (POC) nucleic acid detection technologies are poised to aid gold-standard technologies in controlling the COVID-19 pandemic, yet shortcomings in the capability to perform critically needed complex detection—such as multiplexed detection for viral variant surveillance—may limit their widespread adoption. Herein, we developed a robust multiplexed clustered regularly interspaced short palindromic repeats (CRISPR)-based detection using LwaCas13a and PsmCas13b to simultaneously diagnose severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection and pinpoint the causative SARS-CoV-2 variant of concern (VOC)—including globally dominant VOCs Delta (B.1.617.2) and Omicron (B.1.1.529)—all the while maintaining high levels of accuracy upon the detection of multiple SARS-CoV-2 gene targets. The platform has several attributes suitable for POC use: premixed, freeze-dried reagents for easy use and storage; convenient direct-to-eye or smartphone-based readouts; and a one-pot variant of the multiplexed detection. To reduce reliance on proprietary reagents and enable sustainable use of such a technology in low- and middle-income countries, we locally produced and formulated our own recombinase polymerase amplification reaction and demonstrated its equivalent efficiency to commercial counterparts. Our tool—CRISPR-based detection for simultaneous COVID-19 diagnosis and variant surveillance that can be locally manufactured—may enable sustainable use of CRISPR diagnostics technologies for COVID-19 and other diseases in POC settings. |
Issue Date: | 11-Nov-2022 |
Date of Acceptance: | 1-Nov-2022 |
URI: | http://hdl.handle.net/10044/1/100953 |
DOI: | 10.1089/crispr.2022.0048 |
ISSN: | 2573-1599 |
Publisher: | Mary Ann Liebert |
Start Page: | 1 |
End Page: | 17 |
Journal / Book Title: | The CRISPR Journal |
Volume: | 6 |
Issue: | 2 |
Copyright Statement: | Copyright 2022, Mary Ann Liebert, Inc., publishers. This is the accepted version of the following article: A Multiplexed Cas13-Based Assay with Point-of-Care Attributes for Simultaneous COVID-19 Diagnosis and Variant Surveillance, which has now been formally published in final form at The CRISPR Journal at https://doi.org/10.1089/crispr.2022.0048 This original submission version of the article may be used for non-commercial purposes in accordance with the Mary Ann Liebert, Inc., publishers’ self-archiving terms and conditions. |
Publication Status: | Published online |
Online Publication Date: | 2022-11-11 |
Appears in Collections: | Department of Infectious Diseases Faculty of Medicine Imperial College London COVID-19 |