Streamlined inactivation, amplification, and Cas13-based detection of SARS-CoV-2.
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| Abstract | The COVID-19 pandemic has highlighted that new diagnostic technologies are essential for controlling disease transmission. Here, we develop SHINE (Streamlined Highlighting of Infections to Navigate Epidemics), a sensitive and specific diagnostic tool that can detect SARS-CoV-2 RNA from unextracted samples. We identify the optimal conditions to allow RPA-based amplification and Cas13-based detection to occur in a single step, simplifying assay preparation and reducing run-time. We improve HUDSON to rapidly inactivate viruses in nasopharyngeal swabs and saliva in 10 min. SHINE's results can be visualized with an in-tube fluorescent readout - reducing contamination risk as amplification reaction tubes remain sealed - and interpreted by a companion smartphone application. We validate SHINE on 50 nasopharyngeal patient samples, demonstrating 90% sensitivity and 100% specificity compared to RT-qPCR with a sample-to-answer time of 50 min. SHINE has the potential to be used outside of hospitals and clinical laboratories, greatly enhancing diagnostic capabilities. |
| Year of Publication | 2020
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| Journal | Nat Commun
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| Volume | 11
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| Issue | 1
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| Pages | 5921
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| Date Published | 2020 11 20
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| ISSN | 2041-1723
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| DOI | 10.1038/s41467-020-19097-x
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| PubMed ID | 33219225
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| PubMed Central ID | PMC7680145
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| Grant list | T32 GM007753 / GM / NIGMS NIH HHS / United States
HHMI / Howard Hughes Medical Institute / United States
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