Field-deployable viral diagnostics using CRISPR-Cas13

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Myhrvold, Cameron
Freije, Catherine A.
Gootenberg, Jonathan S.
Abudayyeh, Omar O.
Metsky, Hayden C.
Durbin, Ann F.
Kellner, Max J.
Tan, Amanda L.
Paul, Lauren M.
Parham, Leda A.
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Amer Assoc Advancement Science
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 DENVdetection 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.
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Science. Washington: Amer Assoc Advancement Science, v. 360, n. 6387, p. 444-448, 2018.