Streamlined inactivation, amplification, and Cas13-based detection of SARS-CoV-2
Jon Arizti-Sanz, Catherine A. Freije, Alexandra C. Stanton, Brittany A. Petros, Chloe K. Boehm, Sameed Siddiqui, Bennett M. Shaw, Gordon Adams, Tinna-Solveig F. Kosoko-Thoroddsen, Molly E. Kemball, Jessica N. Uwanibe, Fehintola V. Ajogbasile, Philomena E. Eromon, Robin Gross, Loni Wronka, Katie Caviness, Lisa E. Hensley, Nicholas H. Bergman, Bronwyn L. MacInnis, Christian T. Happi, Jacob E. Lemieux, Pardis C. Sabeti & Cameron Myhrvold
Nature Communications volume 11, Article number: 5921 (2020)
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.
