Clustered regularly interspaced short palindromic repeat (CRISPR/Cas) proteins can be designed to bind specified DNA and RNA sequences and hold great promise for the accurate detection of nucleic acids for diagnostics. We integrated commercially available reagents into a CRISPR/Cas9based lateral flow assay that can detect severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) sequences with single-base specificity. This approach requires minimal equipment and represents a simplified platform for field-based deployment. We also developed a rapid, multiplex fluorescence CRISPR/Cas9 nuclease cleavage assay capable of detecting and differentiating SARS-CoV-2, influenza A and B, and respiratory syncytial virus in a single reaction. Our findings provide proof-of-principle for CRISPR/Cas9 point-of-care diagnosis as well as a scalable fluorescent platform for identifying respiratory viral pathogens with overlapping symptomology.
Bibliographical noteFunding Information:
This research was funded by University of Minnesota Rapid COVID grants from the Medical School and Institute of Engineering in Medicine (M.J.O.). M.J.O. is also supported by a Saint Baldrick?s Foundation Scholar Award.
Funding: This research was funded by University of Minnesota Rapid COVID grants from the Medical School and Institute of Engineering in Medicine (M.J.O.). M.J.O. is also supported by a Saint Baldrick’s Foundation Scholar Award.
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- Lateral flow assay