Field-deployable viral diagnostics using CRISPR-Cas13.
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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. |
Year of Publication | 2018
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Journal | Science
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Volume | 360
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Issue | 6387
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Pages | 444-448
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Date Published | 2018 04 27
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ISSN | 1095-9203
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DOI | 10.1126/science.aas8836
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PubMed ID | 29700266
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PubMed Central ID | PMC6197056
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Grant list | R01 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
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