Development of a Rapid and Sensitive CasRx-Based Diagnostic Assay for SARS-CoV‑2

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.