• Keith Pardee, Alexander A Green, Melissa K Takahashi, Dana Braff, Guillaume Lambert, Jeong Wook Lee, Tom Ferrante, Duo Ma, Nina Donghia, Melina Fan, Nichole M Daringer, Irene Bosch, Dawn M Dudley, David H O'Connor, Lee Gehrke, and James J Collins.
• Leslie Dan Faculty of Pharmacy, University of Toronto, Toronto, ON M5S 3M2, Canada.
• Cell. 2016 May 19; 165 (5): 1255-1266.

AbstractThe recent Zika virus outbreak highlights the need for low-cost diagnostics that can be rapidly developed for distribution and use in pandemic regions. Here, we report a pipeline for the rapid design, assembly, and validation of cell-free, paper-based sensors for the detection of the Zika virus RNA genome. By linking isothermal RNA amplification to toehold switch RNA sensors, we detect clinically relevant concentrations of Zika virus sequences and demonstrate specificity against closely related Dengue virus sequences. When coupled with a novel CRISPR/Cas9-based module, our sensors can discriminate between viral strains with single-base resolution. We successfully demonstrate a simple, field-ready sample-processing workflow and detect Zika virus from the plasma of a viremic macaque. Our freeze-dried biomolecular platform resolves important practical limitations to the deployment of molecular diagnostics in the field and demonstrates how synthetic biology can be used to develop diagnostic tools for confronting global health crises. PAPERCLIP. Copyright © 2016 Elsevier Inc. All rights reserved.

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