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Integrated sample inactivation, amplification, and Cas13-based detection of SARS-CoV-2

Arizti-Sanz, J.; Freije, C. A.; Stanton, A. C.; Boehm, C. K.; Petros, B. A.; Siddiqui, S.; Shaw, B. M.; Adams, G.; Kosoko-Thoroddsen, T.-S. F.; Kemball, M. E.; Gross, R.; Wronka, L.; Caviness, K.; Hensley, L. E.; Bergman, N. H.; MacInnis, B. L.; Lemieux, J. E.; Sabeti, P. C.; Myhrvold, C.

2020-05-28 bioengineering
10.1101/2020.05.28.119131 bioRxiv
Show abstract

The COVID-19 pandemic has highlighted that new diagnostic technologies are essential for controlling disease transmission. Here, we develop SHINE (SHERLOCK and HUDSON Integration to Navigate Epidemics), a sensitive and specific integrated diagnostic tool that can detect SARS-CoV-2 RNA from unextracted samples. We combine the steps of SHERLOCK into a single-step reaction and optimize HUDSON to accelerate viral inactivation in nasopharyngeal swabs and saliva. SHINEs 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-PCR with a sample-to-answer time of 50 minutes. SHINE has the potential to be used outside of hospitals and clinical laboratories, greatly enhancing diagnostic capabilities.

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