Field-deployable viral diagnostics using CRISPR-Cas13.

TitleField-deployable viral diagnostics using CRISPR-Cas13.
Publication TypeJournal Article
Year of Publication2018
AuthorsMyhrvold, C, Freije, CA, Gootenberg, JS, Abudayyeh, OO, Metsky, HC, Durbin, AF, Kellner, MJ, Tan, AL, Paul, LM, Parham, LA, Garcia, KF, Barnes, KG, Chak, B, Mondini, A, Nogueira, ML, Isern, S, Michael, SF, Lorenzana, I, Yozwiak, NL, MacInnis, BL, Bosch, I, Gehrke, L, Zhang, F, Sabeti, PC
JournalScience
Volume360
Issue6387
Pagination444-448
Date Published2018 04 27
ISSN1095-9203
KeywordsAdaptation, Physiological, Bacterial Proteins, CRISPR-Associated Proteins, Dengue, Dengue Virus, Endonucleases, Enzyme Assays, Humans, Microcephaly, Polymorphism, Single Nucleotide, RNA, Viral, Zika Virus, Zika Virus Infection
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.

DOI10.1126/science.aas8836
Alternate JournalScience
PubMed ID29700266
PubMed Central IDPMC6197056
Grant ListR01 AI099210 / AI / NIAID NIH HHS / United States
R01 MH110049 / MH / NIMH NIH HHS / United States
DP1 HL141201 / HL / NHLBI NIH HHS / United States
R33 AI100190 / AI / NIAID NIH HHS / United States
R01 HG009761 / HG / NHGRI NIH HHS / United States
U19 AI110818 / AI / NIAID NIH HHS / United States
R21 AI100190 / AI / NIAID NIH HHS / United States
F30 CA210382 / CA / NCI NIH HHS / United States