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2023 |
A Multiplexed Cas13-Based Assay with Point-of-Care Attributes for Simultaneous COVID-19 Diagnosis and Variant Surveillance |
Maturada Patchsung,Aimorn Homchan,Kanokpol Aphicho,Surased Suraritdechachai,Thanyapat Wanitchanon,Archiraya Pattama,Khomkrit Sappakhaw,Piyachat Meesawat,Thanakrit Wongsatit,Artittaya Athipanyasilp,Krittapas Jantarug,Niracha Athipanyasilp,Juthamas Buahom,Supapat Visanpattanasin,Nootaree Niljianskul,Pimchai Chaiyen,Ruchanok Tinikul,Nuanjun Wichukchinda,Surakameth Mahasirimongkol,Rujipas Sirijatuphat,Nasikarn Angkasekwinai,Michael A Crone,Paul S Freemont,Julia Joung,Alim Ladha,Omar Abudayyeh,Jonathan Gootenberg,Feng Zhang,Claire Chewapreecha,Sittinan Chanarat,Navin Horthongkham,Danaya Pakotiprapha,Chayasith Uttamapinant |
The CRISPR Journal |
36367987 |
10.1089/crispr.2022.0048 |
A Multiplexed Cas13-Based Assay with Point-of-Care Attributes for Simultaneous COVID-19 Diagnosis and Variant Surveillance
Author(s):
Maturada Patchsung,Aimorn Homchan,Kanokpol Aphicho,Surased Suraritdechachai,Thanyapat Wanitchanon,Archiraya Pattama,Khomkrit Sappakhaw,Piyachat Meesawat,Thanakrit Wongsatit,Artittaya Athipanyasilp,Krittapas Jantarug,Niracha Athipanyasilp,Juthamas Buahom,Supapat Visanpattanasin,Nootaree Niljianskul,Pimchai Chaiyen,Ruchanok Tinikul,Nuanjun Wichukchinda,Surakameth Mahasirimongkol,Rujipas Sirijatuphat,Nasikarn Angkasekwinai,Michael A Crone,Paul S Freemont,Julia Joung,Alim Ladha,Omar Abudayyeh,Jonathan Gootenberg,Feng Zhang,Claire Chewapreecha,Sittinan Chanarat,Navin Horthongkham,Danaya Pakotiprapha,Chayasith Uttamapinant
Journal:
The CRISPR Journal
Year:
2023
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.
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