Field-deployable viral diagnostics using CRISPR-Cas13

Citation:

Cameron Myhrvold*†, Catherine A Freije*†, Jonathan S Gootenberg, Omar O Abudayyeh, Hayden C Metsky, Ann F Durbin, Max J Kellner, Amanda L Tan, Lauren M Paul, Leda A Parham, Kimberly F Garcia, Kayla G Barnes, Bridget Chak, Adriano Mondini, Mauricio L Nogueira, Sharon Isern, Scott F Michael, Ivette Lorenzana, Nathan L Yozwiak, Bronwyn L MacInnis, Irene Bosch, Lee Gehrke, Feng Zhang, and Pardis C Sabeti†. 2018. “Field-deployable viral diagnostics using CRISPR-Cas13.” Science, 360, 6387, Pp. 444-448.

ISSN:

1095-9203

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.

DOI:

10.1126/science.aas8836

Alternate Journal:

Science
Last updated on 07/30/2020