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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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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