tetano
Editor, Senior Moderator
CRISPR J
. 2022 Nov 11.
doi: 10.1089/crispr.2022.0048. Online ahead of print.
A Multiplexed Cas13-Based Assay with Point-of-Care Attributes for Simultaneous COVID-19 Diagnosis and Variant Surveillance
Maturada Patchsung[SUP] 1 [/SUP], Aimorn Homchan[SUP] 1 2 [/SUP], Kanokpol Aphicho[SUP] 1 [/SUP], Surased Suraritdechachai[SUP] 1 [/SUP], Thanyapat Wanitchanon[SUP] 1 3 [/SUP], Archiraya Pattama[SUP] 4 [/SUP], Khomkrit Sappakhaw[SUP] 1 [/SUP], Piyachat Meesawat[SUP] 1 [/SUP], Thanakrit Wongsatit[SUP] 1 [/SUP], Artittaya Athipanyasilp[SUP] 1 4 [/SUP], Krittapas Jantarug[SUP] 1 [/SUP], Niracha Athipanyasilp[SUP] 4 [/SUP], Juthamas Buahom[SUP] 4 [/SUP], Supapat Visanpattanasin[SUP] 2 [/SUP], Nootaree Niljianskul[SUP] 5 [/SUP], Pimchai Chaiyen[SUP] 1 [/SUP], Ruchanok Tinikul[SUP] 2 [/SUP], Nuanjun Wichukchinda[SUP] 3 [/SUP], Surakameth Mahasirimongkol[SUP] 3 [/SUP], Rujipas Sirijatuphat[SUP] 6 [/SUP], Nasikarn Angkasekwinai[SUP] 6 [/SUP], Michael A Crone[SUP] 7 8 9 [/SUP], Paul S Freemont[SUP] 7 8 9 [/SUP], Julia Joung[SUP] 10 11 12 13 14 [/SUP], Alim Ladha[SUP] 10 11 12 13 14 [/SUP], Omar Abudayyeh[SUP] 12 [/SUP], Jonathan Gootenberg[SUP] 12 [/SUP], Feng Zhang[SUP] 10 11 12 13 14 [/SUP], Claire Chewapreecha[SUP] 15 16 [/SUP], Sittinan Chanarat[SUP] 2 [/SUP], Navin Horthongkham[SUP] 4 [/SUP], Danaya Pakotiprapha[SUP] 2 [/SUP], Chayasith Uttamapinant[SUP] 1 [/SUP]
Affiliations
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.
. 2022 Nov 11.
doi: 10.1089/crispr.2022.0048. Online ahead of print.
A Multiplexed Cas13-Based Assay with Point-of-Care Attributes for Simultaneous COVID-19 Diagnosis and Variant Surveillance
Maturada Patchsung[SUP] 1 [/SUP], Aimorn Homchan[SUP] 1 2 [/SUP], Kanokpol Aphicho[SUP] 1 [/SUP], Surased Suraritdechachai[SUP] 1 [/SUP], Thanyapat Wanitchanon[SUP] 1 3 [/SUP], Archiraya Pattama[SUP] 4 [/SUP], Khomkrit Sappakhaw[SUP] 1 [/SUP], Piyachat Meesawat[SUP] 1 [/SUP], Thanakrit Wongsatit[SUP] 1 [/SUP], Artittaya Athipanyasilp[SUP] 1 4 [/SUP], Krittapas Jantarug[SUP] 1 [/SUP], Niracha Athipanyasilp[SUP] 4 [/SUP], Juthamas Buahom[SUP] 4 [/SUP], Supapat Visanpattanasin[SUP] 2 [/SUP], Nootaree Niljianskul[SUP] 5 [/SUP], Pimchai Chaiyen[SUP] 1 [/SUP], Ruchanok Tinikul[SUP] 2 [/SUP], Nuanjun Wichukchinda[SUP] 3 [/SUP], Surakameth Mahasirimongkol[SUP] 3 [/SUP], Rujipas Sirijatuphat[SUP] 6 [/SUP], Nasikarn Angkasekwinai[SUP] 6 [/SUP], Michael A Crone[SUP] 7 8 9 [/SUP], Paul S Freemont[SUP] 7 8 9 [/SUP], Julia Joung[SUP] 10 11 12 13 14 [/SUP], Alim Ladha[SUP] 10 11 12 13 14 [/SUP], Omar Abudayyeh[SUP] 12 [/SUP], Jonathan Gootenberg[SUP] 12 [/SUP], Feng Zhang[SUP] 10 11 12 13 14 [/SUP], Claire Chewapreecha[SUP] 15 16 [/SUP], Sittinan Chanarat[SUP] 2 [/SUP], Navin Horthongkham[SUP] 4 [/SUP], Danaya Pakotiprapha[SUP] 2 [/SUP], Chayasith Uttamapinant[SUP] 1 [/SUP]
Affiliations
- PMID: 36367987
- DOI: 10.1089/crispr.2022.0048
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.