A multiplexed Cas13-based assay with point-of-care attributes for simultaneous COVID-19 diagnosis and variant surveillance
File(s)elife-MS-Fig-SI-V7.docx (11.88 MB)
Accepted version
Author(s)
Patchsung, Maturada
Homchan, Aimorn
Aphicho, Kanokpol
Suraritdechachai, Surased
Wanitchanon, Thanyapat
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.
Date Issued
2022-11-11
Date Acceptance
2022-11-01
Citation
The CRISPR Journal, 2022, 6 (2), pp.1-17
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.
Identifier
https://www.webofscience.com/api/gateway?GWVersion=2&SrcApp=PARTNER_APP&SrcAuth=LinksAMR&KeyUT=WOS:000884340600001&DestLinkType=FullRecord&DestApp=ALL_WOS&UsrCustomerID=a2bf6146997ec60c407a63945d4e92bb
Subjects
DNA
Genetics & Heredity
Life Sciences & Biomedicine
NUCLEIC-ACID DETECTION
Science & Technology
Publication Status
Published online
Date Publish Online
2022-11-11