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Deployable CRISPR-Cas13a diagnostic tools to detect and report Ebola and Lassa virus cases in real-time

Article scientifique 2020 Anglais

Résumé

Recent outbreaks of viral hemorrhagic fevers (VHFs), including Ebola virus disease (EVD) and Lassa fever (LF), highlight the urgent need for sensitive, deployable tests to diagnose these devastating human diseases. Here we develop CRISPR-Cas13a-based (SHERLOCK) diagnostics targeting Ebola virus (EBOV) and Lassa virus (LASV), with both fluorescent and lateral flow readouts. We demonstrate on laboratory and clinical samples the sensitivity of these assays and the capacity of the SHERLOCK platform to handle virus-specific diagnostic challenges. We perform safety testing to demonstrate the efficacy of our HUDSON protocol in heat-inactivating VHF viruses before SHERLOCK testing, eliminating the need for an extraction. We develop a user-friendly protocol and mobile application (HandLens) to report results, facilitating SHERLOCK's use in endemic regions. Finally, we successfully deploy our tests in Sierra Leone and Nigeria in response to recent outbreaks.

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Barnes, K., Lachenauer, A., Nitido, A., Siddiqui, S., Gross, R., Beitzel, B., Siddle, K., Freije, C., Dighero-Kemp, B., Mehta, S., Carter, A., Uwanibe, J., Ajogbasile, F., Olumade, T., Odia, I., Sandi, J., Momoh, M., Metsky, H., Boehm, C., Lin, A., Kemball, M., Park, D., Branco, L., Boisen, M., Sullivan, B., Amare, M., Tiamiyu, A., Parker, Z., Iroezindu, M., Grant, D., Modjarrad, K., Myhrvold, C., Garry, R., Palacios, G., Hensley, L., Schaffner, S., Happi, C., Colubri, A., Sabeti, P. (2020). Deployable CRISPR-Cas13a diagnostic tools to detect and report Ebola and Lassa virus cases in real-time. https://doi.org/10.1038/s41467-020-17994-9

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Auteur(s)

Kayla G. Barnes Anna E. Lachenauer Adam D. Nitido Sameed M. Siddiqui Robin Gross Brett Beitzel Katherine J. Siddle Catherine A. Freije Bonnie Dighero-Kemp Samar Mehta Amber Carter Jessica N. Uwanibe Fehintola V. Ajogbasile Testimony Olumade Ikponmwosa Odia John Demby Sandi Mambu Momoh Hayden C. Metsky Chloe K. Boehm Aaron E. Lin Molly Kemball Daniel J. Park Luis M. Branco Matthew L. Boisen Brian M. Sullivan Mihret F. Amare Abdulwasiu Bolaji Tiamiyu Zahra Parker Michael Iroezindu Donald S. Grant Kayvon Modjarrad Cameron Myhrvold Robert F. Garry Gustavo Palacios Lisa E. Hensley S. F. Schaffner Christian Happi Andrés Colubri Pardis C. Sabeti

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