Development of a Rapid and Sensitive CasRx-Based Diagnostic Assay for SARS-CoV-2.
Daniel J Brogan, Duverney Chaverra-Rodriguez, Calvin P Lin, Andrea L Smidler, Ting Yang, Lenissa M Alcantara, Igor Antoshechkin, Junru Liu, Robyn R Raban, Pedro Belda-Ferre, Rob Knight, Elizabeth A Komives, Omar S Akbari
ACS sensors · 2021-Nov-26 · PMID 34714054 · DOI 10.1021/acssensors.1c01088Abstract
The development of an extensive toolkit for potential point-of-care diagnostics that is expeditiously adaptable to new emerging pathogens is of critical public health importance. Recently, a number of novel CRISPR-based diagnostics have been developed to detect SARS-CoV-2. Herein, we outline the development of an alternative CRISPR nucleic acid diagnostic utilizing a Cas13d ribonuclease derived from Ruminococcus flavefaciens XPD3002 (CasRx) to detect SARS-CoV-2, an approach we term SENSR (sensitive enzymatic nucleic acid sequence reporter) that can detect attomolar concentrations of SARS-CoV-2. We demonstrate 100% sensitivity in patient-derived samples by lateral flow and fluorescence readout with a detection limit of 45 copy/μL. This technology expands the available nucleic acid diagnostic toolkit, which can be adapted to combat future pandemics.
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- laboratory
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