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 Alcantara, Igor Antoshechkin, Junru Liu, Robyn Raban, Pedro Belda‐Ferre, Rob Knight, Elizabeth A. Komives, Omar S. Akbari
Abstract
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.