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

Antibodies, Viral Clustered Regularly Interspaced Short Palindromic Repeats / genetics Ebolavirus / genetics Hemorrhagic Fever, Ebola / diagnosis Lassa Fever / diagnosis Lassa virus / genetics

Journal

Nature communications
ISSN: 2041-1723
Titre abrégé: Nat Commun
Pays: England
ID NLM: 101528555

Informations de publication

Date de publication:
17 08 2020
Historique:
received: 20 03 2020
accepted: 28 07 2020
entrez: 19 8 2020
pubmed: 19 8 2020
medline: 9 9 2020
Statut: epublish

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.

Identifiants

DOI: 10.1038/s41467-020-17994-9 PMID: 32807807 PMC: PMC7431545
pubmed: 32807807
doi: 10.1038/s41467-020-17994-9
pii: 10.1038/s41467-020-17994-9
pmc: PMC7431545
doi:

Substances chimiques

Antibodies, Viral 0

Types de publication

Journal Article Research Support, N.I.H., Extramural Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S.

Langues

eng

Sous-ensembles de citation

IM

Pagination

4131

Subventions

Organisme : NIAID NIH HHS
ID : R01 AI114855
Pays : United States
Organisme : Howard Hughes Medical Institute
Pays : United States
Organisme : NIAID NIH HHS
ID : R01 AI132223
Pays : United States
Organisme : FIC NIH HHS
ID : K01 TW010853
Pays : United States
Organisme : NIAID NIH HHS
ID : R01 AI132244
Pays : United States
Organisme : NIAID NIH HHS
ID : U01 AI151812
Pays : United States
Organisme : NHGRI NIH HHS
ID : U54 HG007480
Pays : United States
Organisme : NIAID NIH HHS
ID : U19 AI110818
Pays : United States

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Auteurs

Kayla G Barnes (KG)

Broad Institute of MIT and Harvard, Cambridge, Massachusetts, USA. kbarnes@broadinstitute.org.
Department of Immunology and Infectious Diseases, Harvard T.H. Chan School of Public Health, Harvard University, Boston, Massachusetts, USA. kbarnes@broadinstitute.org.
MRC-University of Glasgow Centre for Virus Research, Glasgow, UK. kbarnes@broadinstitute.org.
ORCID: 0000-0002-8291-4388

Anna E Lachenauer (AE)

Broad Institute of MIT and Harvard, Cambridge, Massachusetts, USA. annalach@stanford.edu.
Stanford University School of Medicine, Stanford, California, USA. annalach@stanford.edu.
ORCID: 0000-0002-2930-9541

Adam Nitido (A)

Ragon Institute of MGH, MIT, and Harvard, Cambridge, Massachusetts, 02139, USA.
Ph.D. Program in Virology, Division of Medical Sciences, Harvard Medical School, Boston, Massachusetts, 02115, USA.
ORCID: 0000-0001-5679-838X

Sameed Siddiqui (S)

Broad Institute of MIT and Harvard, Cambridge, Massachusetts, USA.
Computational and Systems Biology, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA.

Robin Gross (R)

Integrated Research Facility, Division of Clinical Research, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Frederick, Maryland, USA.

Brett Beitzel (B)

Center for Genome Sciences, The United States Army Medical Research Institute for Infectious Disease, 1425 Porter Street, Fort Detrick, Maryland, 21702, USA.
ORCID: 0000-0002-3798-6411

Katherine J Siddle (KJ)

Broad Institute of MIT and Harvard, Cambridge, Massachusetts, USA.
Center for Systems Biology, Department of Organismic and Evolutionary Biology, Harvard University, Cambridge, Massachusetts, USA.

Catherine A Freije (CA)

Broad Institute of MIT and Harvard, Cambridge, Massachusetts, USA.
Ph.D. Program in Virology, Division of Medical Sciences, Harvard Medical School, Boston, Massachusetts, 02115, USA.
ORCID: 0000-0002-1939-3102

Bonnie Dighero-Kemp (B)

Integrated Research Facility, Division of Clinical Research, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Frederick, Maryland, USA.

Samar B Mehta (SB)

Broad Institute of MIT and Harvard, Cambridge, Massachusetts, USA.
Division of Infectious Diseases, Beth Israel Deaconess Medical Center, Boston, Massachusetts, USA.

Amber Carter (A)

Broad Institute of MIT and Harvard, Cambridge, Massachusetts, USA.

Jessica Uwanibe (J)

African Center of Excellence for Genomics of Infectious Disease (ACEGID), Redeemer's University, Ede, Osun State, Nigeria.
Department of Biological Sciences, College of Natural Sciences, Redeemer's University, Ede, Osun State, Nigeria.

Fehintola Ajogbasile (F)

African Center of Excellence for Genomics of Infectious Disease (ACEGID), Redeemer's University, Ede, Osun State, Nigeria.
Department of Biological Sciences, College of Natural Sciences, Redeemer's University, Ede, Osun State, Nigeria.

Testimony Olumade (T)

African Center of Excellence for Genomics of Infectious Disease (ACEGID), Redeemer's University, Ede, Osun State, Nigeria.
Department of Biological Sciences, College of Natural Sciences, Redeemer's University, Ede, Osun State, Nigeria.
ORCID: 0000-0002-5046-153X

Ikponmwosa Odia (I)

Institute of Lassa Fever Research and Control, Irrua Specialist Teaching Hospital, Irrua, Edo State, Nigeria.

John Demby Sandi (JD)

African Center of Excellence for Genomics of Infectious Disease (ACEGID), Redeemer's University, Ede, Osun State, Nigeria.
Viral Hemorrhagic Fever Program, Kenema Government Hospital, Kenema, Sierra Leone.

Mambu Momoh (M)

African Center of Excellence for Genomics of Infectious Disease (ACEGID), Redeemer's University, Ede, Osun State, Nigeria.
Viral Hemorrhagic Fever Program, Kenema Government Hospital, Kenema, Sierra Leone.

Hayden C Metsky (HC)

Broad Institute of MIT and Harvard, Cambridge, Massachusetts, USA.
Department of Electrical Engineering and Computer Science, MIT, Cambridge, Massachusetts, 02139, USA.

Chloe K Boehm (CK)

Broad Institute of MIT and Harvard, Cambridge, Massachusetts, USA.
ORCID: 0000-0002-6136-3257

Aaron E Lin (AE)

Broad Institute of MIT and Harvard, Cambridge, Massachusetts, USA.
Ph.D. Program in Virology, Division of Medical Sciences, Harvard Medical School, Boston, Massachusetts, 02115, USA.
ORCID: 0000-0001-7400-4125

Molly Kemball (M)

Broad Institute of MIT and Harvard, Cambridge, Massachusetts, USA.
Center for Systems Biology, Department of Organismic and Evolutionary Biology, Harvard University, Cambridge, Massachusetts, USA.

Daniel J Park (DJ)

Broad Institute of MIT and Harvard, Cambridge, Massachusetts, USA.
ORCID: 0000-0001-7226-7781

Luis Branco (L)

Zalgen Labs, Germantown, Maryland, USA.

Matt Boisen (M)

Zalgen Labs, Germantown, Maryland, USA.

Brian Sullivan (B)

Department of Immunology and Microbial Science, The Scripps Research Institute, La Jolla, California, USA.
ORCID: 0000-0003-4277-6037

Mihret F Amare (MF)

Emerging Infectious Diseases Branch, Walter Reed Army Institute of Research, Silver Spring, Maryland, USA.
Henry M. Jackson Foundation for the Advancement of Military Medicine, Bethesda, Maryland, USA.

Abdulwasiu B Tiamiyu (AB)

Emerging Infectious Diseases Branch, Walter Reed Army Institute of Research, Silver Spring, Maryland, USA.
Henry M. Jackson Foundation Medical Research International, Abuja, Nigeria.

Zahra F Parker (ZF)

Emerging Infectious Diseases Branch, Walter Reed Army Institute of Research, Silver Spring, Maryland, USA.
Henry M. Jackson Foundation for the Advancement of Military Medicine, Bethesda, Maryland, USA.

Michael Iroezindu (M)

Emerging Infectious Diseases Branch, Walter Reed Army Institute of Research, Silver Spring, Maryland, USA.
Henry M. Jackson Foundation Medical Research International, Abuja, Nigeria.

Donald S Grant (DS)

Viral Hemorrhagic Fever Program, Kenema Government Hospital, Kenema, Sierra Leone.
Ministry of Health and Sanitation, Freetown, Sierra Leone.
ORCID: 0000-0002-4329-0795

Kayvon Modjarrad (K)

Emerging Infectious Diseases Branch, Walter Reed Army Institute of Research, Silver Spring, Maryland, USA.

Cameron Myhrvold (C)

Broad Institute of MIT and Harvard, Cambridge, Massachusetts, USA.
Center for Systems Biology, Department of Organismic and Evolutionary Biology, Harvard University, Cambridge, Massachusetts, USA.

Robert F Garry (RF)

Zalgen Labs, Germantown, Maryland, USA.
Tulane University School of Medicine, New Orleans, Los Angeles, 70112, USA.

Gustavo Palacios (G)

Center for Genome Sciences, The United States Army Medical Research Institute for Infectious Disease, 1425 Porter Street, Fort Detrick, Maryland, 21702, USA.

Lisa E Hensley (LE)

Integrated Research Facility, Division of Clinical Research, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Frederick, Maryland, USA.

Stephen F Schaffner (SF)

Broad Institute of MIT and Harvard, Cambridge, Massachusetts, USA.
Department of Immunology and Infectious Diseases, Harvard T.H. Chan School of Public Health, Harvard University, Boston, Massachusetts, USA.
Center for Systems Biology, Department of Organismic and Evolutionary Biology, Harvard University, Cambridge, Massachusetts, USA.
ORCID: 0000-0001-6699-3568

Christian T Happi (CT)

Department of Immunology and Infectious Diseases, Harvard T.H. Chan School of Public Health, Harvard University, Boston, Massachusetts, USA.
African Center of Excellence for Genomics of Infectious Disease (ACEGID), Redeemer's University, Ede, Osun State, Nigeria.
Department of Biological Sciences, College of Natural Sciences, Redeemer's University, Ede, Osun State, Nigeria.
Institute of Lassa Fever Research and Control, Irrua Specialist Teaching Hospital, Irrua, Edo State, Nigeria.

Andres Colubri (A)

Broad Institute of MIT and Harvard, Cambridge, Massachusetts, USA.
Center for Systems Biology, Department of Organismic and Evolutionary Biology, Harvard University, Cambridge, Massachusetts, USA.

Pardis C Sabeti (PC)

Broad Institute of MIT and Harvard, Cambridge, Massachusetts, USA.
Department of Immunology and Infectious Diseases, Harvard T.H. Chan School of Public Health, Harvard University, Boston, Massachusetts, USA.
Center for Systems Biology, Department of Organismic and Evolutionary Biology, Harvard University, Cambridge, Massachusetts, USA.
Howard Hughes Medical Institute, Chevy Chase, Maryland, USA.

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Classifications MeSH

questionsmedicales.fr › Acides aminés, peptides et protéines › Protéines › Globulines › Sérum-globulines › Immunoglobulines › Anticorps › Anticorps antiviraux › Deployable CRISPR-Cas13a diagnostic tools to...
questionsmedicales.fr › Phénomènes génétiques › Structures génétiques › Génome › Composants de génome › Séquences répétées en tandem › Clustered regularly interspaced short palindromic repeats › Deployable CRISPR-Cas13a diagnostic tools to...
questionsmedicales.fr › Virus › Virus à ARN › Virus à ARN de polarité négative › Mononegavirales › Filoviridae › Ebolavirus › Deployable CRISPR-Cas13a diagnostic tools to...
questionsmedicales.fr › Infection › Maladies virales › Infections à virus à ARN › Infections à Mononegavirales › Infections à Filoviridae › Fièvre hémorragique à virus Ebola › Deployable CRISPR-Cas13a diagnostic tools to...
questionsmedicales.fr › Infection › Maladies virales › Infections à virus à ARN › Fièvres hémorragiques virales › Fièvre de Lassa › Deployable CRISPR-Cas13a diagnostic tools to...
questionsmedicales.fr › Virus › Virus à ARN › Virus à ARN de polarité négative › Arenaviridae › Arenavirus › Arénavirus de l'Ancien Monde › Virus de Lassa › Deployable CRISPR-Cas13a diagnostic tools to...