A Multiplexed Cas13-Based Assay with Point-of-Care Attributes for Simultaneous COVID-19 Diagnosis and Variant Surveillance
Issued Date
2023-04-01
Resource Type
ISSN
25731599
eISSN
25731602
Scopus ID
2-s2.0-85158830178
Pubmed ID
36367987
Journal Title
CRISPR Journal
Volume
6
Issue
2
Start Page
99
End Page
115
Rights Holder(s)
SCOPUS
Bibliographic Citation
CRISPR Journal Vol.6 No.2 (2023) , 99-115
Suggested Citation
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 M.A., Freemont P.S., Joung J., Ladha A., Abudayyeh O., Gootenberg J., Zhang F., Chewapreecha C., Chanarat S., Horthongkham N., Pakotiprapha D., Uttamapinant C. A Multiplexed Cas13-Based Assay with Point-of-Care Attributes for Simultaneous COVID-19 Diagnosis and Variant Surveillance. CRISPR Journal Vol.6 No.2 (2023) , 99-115. 115. doi:10.1089/crispr.2022.0048 Retrieved from: https://repository.li.mahidol.ac.th/handle/20.500.14594/82646
Title
A Multiplexed Cas13-Based Assay with Point-of-Care Attributes for Simultaneous COVID-19 Diagnosis and Variant Surveillance
Author(s)
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 M.A.
Freemont P.S.
Joung J.
Ladha A.
Abudayyeh O.
Gootenberg J.
Zhang F.
Chewapreecha C.
Chanarat S.
Horthongkham N.
Pakotiprapha D.
Uttamapinant C.
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 M.A.
Freemont P.S.
Joung J.
Ladha A.
Abudayyeh O.
Gootenberg J.
Zhang F.
Chewapreecha C.
Chanarat S.
Horthongkham N.
Pakotiprapha D.
Uttamapinant C.
Author's Affiliation
Siriraj Hospital
Mahidol Oxford Tropical Medicine Research Unit
PTT Public Company Limited
Vidyasirimedhi Institute of Science and Technology
McGovern Institute
Massachusetts Institute of Technology
Thailand Ministry of Public Health
Imperial College London
Mahidol University
Howard Hughes Medical Institute
Wellcome Sanger Institute
Broad Institute
Mahidol Oxford Tropical Medicine Research Unit
PTT Public Company Limited
Vidyasirimedhi Institute of Science and Technology
McGovern Institute
Massachusetts Institute of Technology
Thailand Ministry of Public Health
Imperial College London
Mahidol University
Howard Hughes Medical Institute
Wellcome Sanger Institute
Broad Institute
Other Contributor(s)
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.