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

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

in 45 / 45/77 / 45/90 / 631/208/4041 / 631/326/596 / 692/699/255/2514 / Antibodies, Viral / Applications programs / Clustered Regularly Interspaced Short Palindromic Repeats - genetics / CRISPR / Diagnostic software / Diagnostic systems / Ebola virus / Ebolavirus / Ebolavirus - genetics / Ebolavirus - pathogenicity / Fever / Fluorescence / Hemorrhage / Hemorrhagic Fever, Ebola - diagnosis / Hemorrhagic Fever, Ebola - virology / Humanities and Social Sciences / Lassa fever / Lassa Fever - diagnosis / Lassa Fever - virology / Lassa virus - genetics / Lassa virus - pathogenicity / Mobile computing / multidisciplinary / Outbreaks / Science / Science (multidisciplinary) / Viral diseases / Viruses

2020

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2020

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2020

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Journal Article

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

Sullivan, Brian,

Uwanibe, Jessica,

Dighero-Kemp, Bonnie,

Boehm, Chloe K.,

Siddle, Katherine J.,

Amare, Mihret F.,

Myhrvold, Cameron,

Grant, Donald S.,

Happi, Christian T.,

Gross, Robin,

Lachenauer, Anna E.,

Momoh, Mambu,

Branco, Luis,

Olumade, Testimony,

Ajogbasile, Fehintola,

Schaffner, Stephen F.,

Palacios, Gustavo,

Odia, Ikponmwosa,

Colubri, Andres,

Sabeti, Pardis C.,

Lin, Aaron E.,

Park, Daniel J.,

Beitzel, Brett,

Parker, Zahra F.,

Barnes, Kayla G.,

Iroezindu, Michael,

Carter, Amber,

Sandi, John Demby,

Modjarrad, Kayvon,

Metsky, Hayden C.,

Nitido, Adam,

Freije, Catherine A.,

Mehta, Samar B.,

Kemball, Molly,

Tiamiyu, Abdulwasiu B.,

Boisen, Matt,

Garry, Robert F.,

Hensley, Lisa E.,

Siddiqui, Sameed

2020

Overview

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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Nature Publishing Group UK,Nature Publishing Group,Nature Portfolio

Subject

/ 45/77

/ 45/90

/ Applications programs