Title | Field-deployable viral diagnostics using CRISPR-Cas13. |
Publication Type | Journal Article |
Year of Publication | 2018 |
Authors | Myhrvold, 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 |
Journal | Science |
Volume | 360 |
Issue | 6387 |
Pagination | 444-448 |
Date Published | 2018 Apr 27 |
ISSN | 1095-9203 |
Keywords | Adaptation, 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 | <p>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.</p> |
DOI | 10.1126/science.aas8836 |
Alternate Journal | Science |
PubMed ID | 29700266 |
PubMed Central ID | PMC6197056 |
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 |