Field-deployable viral diagnostics using CRISPR-Cas13

Cameron Myhrvold(Broad Institute), Catherine A. Freije(Broad Institute), Jonathan S. Gootenberg(Broad Institute), Omar O. Abudayyeh(Broad Institute), Hayden C. Metsky(Broad Institute), Ann Durbin(Harvard University), Max J. Kellner(Broad Institute), Amanda L. Tan(Florida Gulf Coast University), Lauren M. Paul(Florida Gulf Coast University), Leda Parham(National Autonomous University of Honduras), Kimberly García(National Autonomous University of Honduras), Kayla G. Barnes(Broad Institute), Bridget Chak(Broad Institute), Adriano Mondini(Centro Universitário de Araraquara), Maurício Lacerda Nogueira(Faculdade de Medicina de São José do Rio Preto), Sharon Isern(Florida Gulf Coast University), Scott F. Michael(Florida Gulf Coast University), Ivette Lorenzana(National Autonomous University of Honduras), Nathan L. Yozwiak(Broad Institute), Bronwyn MacInnis(Broad Institute), Irene Bosch(Icahn School of Medicine at Mount Sinai), Lee Gehrke(Harvard University), Feng Zhang(Broad Institute), Pardis C. Sabeti(Broad Institute)
Science
April 26, 2018
10.1126/science.aas8836
Cited by 1,449Open Access
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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.

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