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Science. 2018 Apr 27;360(6387):444-448. doi: 10.1126/science.aas8836.

Field-deployable viral diagnostics using CRISPR-Cas13.

Myhrvold C1,2, Freije CA1,2,3, Gootenberg JS4,5,6,7,8, Abudayyeh OO4,6,7,8,9, Metsky HC4,10, Durbin AF3,11, Kellner MJ4, Tan AL12, Paul LM12, Parham LA13, Garcia KF13, Barnes KG4,2,14, Chak B4,2, Mondini A15, Nogueira ML16, Isern S12, Michael SF12, Lorenzana I13, Yozwiak NL4,2, MacInnis BL4,14, Bosch I11,17, Gehrke L3,11,18, Zhang F4,6,7,8, Sabeti PC1,2,3,14,19.

Author information

1
Broad Institute of the Massachusetts Institute of Technology (MIT) and Harvard, Cambridge, MA 02142, USA. pardis@broadinstitute.org cmyhrvol@broadinstitute.org cfreije@broadinstitute.org.
2
Center for Systems Biology, Department of Organismal and Evolutionary Biology, Harvard University, Cambridge, MA 02138, USA.
3
Ph.D. Program in Virology, Division of Medical Sciences, Harvard Medical School, Boston, MA 02115, USA.
4
Broad Institute of the Massachusetts Institute of Technology (MIT) and Harvard, Cambridge, MA 02142, USA.
5
Department of Systems Biology, Harvard Medical School, Boston, MA 02115, USA.
6
McGovern Institute for Brain Research, MIT, Cambridge, MA 02139, USA.
7
Department of Brain and Cognitive Science, MIT, Cambridge, MA 02139, USA.
8
Department of Biological Engineering, MIT, Cambridge, MA 02139, USA.
9
Department of Health Sciences and Technology, MIT, Cambridge, MA 02139, USA.
10
Department of Electrical Engineering and Computer Science, MIT, Cambridge, MA 02139, USA.
11
Institute for Medical Engineering and Science, MIT, Cambridge, MA 02139, USA.
12
Department of Biological Sciences, Florida Gulf Coast University, Fort Myers, FL 33965, USA.
13
Centro de Investigaciones Genética, Instituto de Investigacion en Microbiologia, Universidad Nacional Autónoma de Honduras, Tegucigalpa, Honduras.
14
Department of Immunology and Infectious Disease, Harvard School of Public Health, Boston, MA 02115, USA.
15
Araraquara Laboratory of Public Health, School of Pharmaceutical Sciences, São Paulo State University, São Paulo, Brazil.
16
Laboratorio de Pesquisas em Virologia, Faculdade de Medicina de Sao Jose do Rio Preto, São Paulo, Brazil.
17
Department of Medicine, Mount Sinai School of Medicine, New York, NY 10029, USA.
18
Department of Microbiology and Immunobiology, Harvard Medical School, Boston, MA 02115, USA.
19
Howard Hughes Medical Institute (HHMI), Chevy Chase, MD 20815, USA.

Abstract

Mitigating global infectious disease requires diagnostic tools that are sensitive, specific, and rapidly field deployable. In this study, we demonstrate that the Cas13-based SHERLOCK (specific high-sensitivity enzymatic reporter unlocking) platform can detect Zika virus (ZIKV) and dengue virus (DENV) in patient samples at concentrations as low as 1 copy per microliter. We developed HUDSON (heating unextracted diagnostic samples to obliterate nucleases), a protocol that pairs with SHERLOCK for viral detection directly from bodily fluids, enabling instrument-free DENV detection directly from patient samples in <2 hours. We further demonstrate that SHERLOCK can distinguish the four DENV serotypes, as well as region-specific strains of ZIKV from the 2015-2016 pandemic. Finally, we report the rapid (<1 week) design and testing of instrument-free assays to detect clinically relevant viral single-nucleotide polymorphisms.

PMID:
29700266
PMCID:
PMC6197056
DOI:
10.1126/science.aas8836
[Indexed for MEDLINE]
Free PMC Article

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