TY - JOUR
T1 - Deployable CRISPR-Cas13a diagnostic tools to detect and report Ebola and Lassa virus cases in real-time
AU - Barnes, Kayla G.
AU - Lachenauer, Anna E.
AU - Nitido, Adam
AU - Siddiqui, Sameed
AU - Gross, Robin
AU - Beitzel, Brett
AU - Siddle, Katherine J.
AU - Freije, Catherine A.
AU - Dighero-Kemp, Bonnie
AU - Mehta, Samar B.
AU - Carter, Amber
AU - Uwanibe, Jessica
AU - Ajogbasile, Fehintola
AU - Olumade, Testimony
AU - Odia, Ikponmwosa
AU - Sandi, John Demby
AU - Momoh, Mambu
AU - Metsky, Hayden C.
AU - Boehm, Chloe K.
AU - Lin, Aaron E.
AU - Kemball, Molly
AU - Park, Daniel J.
AU - Branco, Luis
AU - Boisen, Matt
AU - Sullivan, Brian
AU - Amare, Mihret F.
AU - Tiamiyu, Abdulwasiu B.
AU - Parker, Zahra F.
AU - Iroezindu, Michael
AU - Grant, Donald S.
AU - Modjarrad, Kayvon
AU - Myhrvold, Cameron
AU - Garry, Robert F.
AU - Palacios, Gustavo
AU - Hensley, Lisa E.
AU - Schaffner, Stephen F.
AU - Happi, Christian T.
AU - Colubri, Andres
AU - Sabeti, Pardis C.
N1 - Funding Information:
Opinions, interpretations, conclusions, and recommendations are those of the authors and are not necessarily endorsed by the United States Army or the National Institute of Health. We thank the staff of the Joint West Africa Research Group, the Walter Reed Army Institute of Research, the Henry M. Jackson Foundation for the Advancement of Military Medicine, and the U.S. Embassy in Abuja, Nigeria, for their collaborative support and our colleagues at the Irrua Specialist Teaching Hospital, Nigeria, and Kenema Government Hospital, Sierra Leone. We thank Dr. Christopher Moxon for his critical edits of the manuscript. This work is supported by grants from the National Institute of Allergy and Infectious Diseases (NIAID), National Institutes of Health (NIH) (U19AI110818 and R01AI114855 to the Broad Institute, P.C.S.), Henry M Jackson Foundation (W81XWH-18-2-0040 to P.C.S.), DARPA (D18AC00006 to the Broad Institute, P.C.S.), the Bill and Melinda Gates Foundation (OPP1192035 to Harvard University, P.C.S.), NIH-Fogarty (K01TW010853 to HTHCSPH, K.G.B.), and P.C.S. is supported by the Howard Hughes Medical Institute.
Publisher Copyright:
© 2020, The Author(s).
PY - 2020/12/1
Y1 - 2020/12/1
N2 - 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.
AB - 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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UR - http://www.scopus.com/inward/citedby.url?scp=85089533250&partnerID=8YFLogxK
U2 - 10.1038/s41467-020-17994-9
DO - 10.1038/s41467-020-17994-9
M3 - Article
C2 - 32807807
AN - SCOPUS:85089533250
SN - 2041-1723
VL - 11
JO - Nature communications
JF - Nature communications
IS - 1
M1 - 4131
ER -