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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 CRISPR-based detection using LwaCas13a and PsmCas13b to simultaneously diagnose 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 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 which 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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Texte intégral: Disponible Collection: Preprints Base de données: medRxiv Sujet Principal: COVID-19 langue: Anglais Année: 2022 Type de document: Preprint

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Texte intégral: Disponible Collection: Preprints Base de données: medRxiv Sujet Principal: COVID-19 langue: Anglais Année: 2022 Type de document: Preprint