4.8 Article

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

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NATURE COMMUNICATIONS
卷 11, 期 1, 页码 -

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NATURE PORTFOLIO
DOI: 10.1038/s41467-020-17994-9

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资金

  1. National Institute of Allergy and Infectious Diseases (NIAID)
  2. National Institutes of Health (NIH) [U19AI110818, R01AI114855]
  3. Henry M Jackson Foundation [W81XWH-18-2-0040]
  4. DARPA [D18AC00006]
  5. Bill and Melinda Gates Foundation [OPP1192035]
  6. NIH-Fogarty [K01TW010853]
  7. Howard Hughes Medical Institute
  8. Bill and Melinda Gates Foundation [OPP1192035] Funding Source: Bill and Melinda Gates Foundation

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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. Outbreaks of viral hemorrhagic fevers highlight the need for sensitive, field-deployable diagnostics. Here the authors present a CRISPR-based SHERLOCK platform with field protocol and mobile app for Ebola and Lassa fever outbreaks.

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