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

被引：108

作者：

Barnes, Kayla G. ^{[1
,2
,3
]}

Lachenauer, Anna E. ^{[1
,4
]}

Nitido, Adam ^{[5
,6
]}

Siddiqui, Sameed ^{[1
,7
]}

Gross, Robin ^{[8
]}

Beitzel, Brett ^{[9
]}

Siddle, Katherine J. ^{[1
,10
]}

Freije, Catherine A. ^{[1
,6
]}

Dighero-Kemp, Bonnie ^{[8
]}

Mehta, Samar B. ^{[1
,11
]}

Carter, Amber ^{[1
]}

Uwanibe, Jessica ^{[12
,13
]}

Ajogbasile, Fehintola ^{[12
,13
]}

Olumade, Testimony ^{[12
,13
]}

Odia, Ikponmwosa ^{[14
]}

Sandi, John Demby ^{[12
,15
]}

Momoh, Mambu ^{[12
,15
]}

Metsky, Hayden C. ^{[1
,16
]}

Boehm, Chloe K. ^{[1
]}

Lin, Aaron E. ^{[1
,6
]}

Kemball, Molly ^{[1
,10
]}

Park, Daniel J. ^{[1
]}

Branco, Luis ^{[17
]}

Boisen, Matt ^{[17
]}

Sullivan, Brian ^{[18
]}

Amare, Mihret F. ^{[19
,20
]}

Tiamiyu, Abdulwasiu B. ^{[19
,21
]}

Parker, Zahra F. ^{[19
,20
]}

Iroezindu, Michael ^{[19
,21
]}

Grant, Donald S. ^{[15
,22
]}

Modjarrad, Kayvon ^{[19
]}

Myhrvold, Cameron ^{[1
,10
]}

Garry, Robert F. ^{[17
,23
]}

Palacios, Gustavo ^{[9
]}

Hensley, Lisa E. ^{[8
]}

Schaffner, Stephen F. ^{[1
,2
,10
]}

Happi, Christian T. ^{[2
,12
,13
,14
]}

Colubri, Andres ^{[1
,10
]}

Sabeti, Pardis C. ^{[1
,2
,10
,24
]}

机构：

[1] Broad Inst MIT & Harvard, Cambridge, MA 02142 USA

[2] Harvard Univ, Dept Immunol & Infect Dis, Harvard TH Chan Sch Publ Hlth, Boston, MA 02115 USA

[3] Univ Glasgow, MRC, Ctr Virus Res, Glasgow, Lanark, Scotland

[4] Stanford Univ, Sch Med, Stanford, CA 94305 USA

[5] Ragon Inst MGH MIT & Harvard, Cambridge, MA 02139 USA

[6] Harvard Med Sch, Div Med Sci, PhD Program Virol, Boston, MA 02115 USA

[7] MIT, Computat & Syst Biol, 77 Massachusetts Ave, Cambridge, MA 02139 USA

[8] NIAID, Integrated Res Facil, Div Clin Res, NIH, Frederick, MD USA

[9] US Army, Ctr Genome Sci, Med Res Inst Infect Dis, 1425 Porter St, Ft Detrick, MD 21702 USA

[10] Harvard Univ, Dept Organism & Evolutionary Biol, Ctr Syst Biol, Cambridge, MA 02138 USA

[11] Beth Israel Deaconess Med Ctr, Div Infect Dis, Boston, MA USA

[12] Redeemers Univ, African Ctr Excellence Genom Infect Dis ACEGID, Ede, Osun State, Nigeria

[13] Redeemers Univ, Coll Nat Sci, Dept Biol Sci, Ede, Osun State, Nigeria

[14] Irrua Specialist Teaching Hosp, Inst Lassa Fever Res & Control, Irrua, Edo State, Nigeria

[15] Kenema Govt Hosp, Viral Hemorrhag Fever Program, Kenema, Sierra Leone

[16] MIT, Dept Elect Engn & Comp Sci, Cambridge, MA 02139 USA

[17] Zalgen Labs, Germantown, MD USA

[18] Scripps Res Inst, Dept Immunol & Microbial Sci, La Jolla, CA USA

[19] Walter Reed Army Inst Res, Emerging Infect Dis Branch, Silver Spring, MD USA

[20] Henry M Jackson Fdn Adv Mil Med, Bethesda, MD USA

[21] Henry M Jackson Fdn Med Res Int, Abuja, Nigeria

[22] Minist Hlth & Sanit, Freetown, Sierra Leone

[23] Tulane Univ, Sch Med, Los Angeles, CA 70112 USA

[24] Howard Hughes Med Inst, Chevy Chase, MD USA

来源：

NATURE COMMUNICATIONS | 2020年 / 11卷 / 01期

基金：

美国国家卫生研究院;

关键词：

NUCLEIC-ACID DETECTION; HEMORRHAGIC-FEVER; TRANSMISSION; EVOLUTION; RISK; CONGO; RNA;

D O I：

10.1038/s41467-020-17994-9

中图分类号：

O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];

学科分类号：

07 ; 0710 ; 09 ;

摘要：

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.

引用

页数：10

共 18 条

[1] Deployable CRISPR-Cas13a diagnostic tools to detect and report Ebola and Lassa virus cases in real-time
Kayla G. Barnes
Anna E. Lachenauer
Adam Nitido
Sameed Siddiqui
Robin Gross
Brett Beitzel
Katherine J. Siddle
Catherine A. Freije
Bonnie Dighero-Kemp
Samar B. Mehta
Amber Carter
Jessica Uwanibe
Fehintola 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 Branco
Matt Boisen
Brian Sullivan
Mihret F. Amare
Abdulwasiu B. Tiamiyu
Zahra F. Parker
Michael Iroezindu
Donald S. Grant
Kayvon Modjarrad
Cameron Myhrvold
Robert F. Garry
Gustavo Palacios
Lisa E. Hensley
Stephen F. Schaffner
Christian T. Happi
Andres Colubri
Pardis C. Sabeti
[J]. Nature Communications, 11
[2] Rapid and Fully Microfluidic Ebola Virus Detection with CRISPR-Cas13a
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