|Publication Type||Journal Article|
|Year of Publication||2016|
|Authors||Pardee, K, Green, AA, Takahashi, MK, Braff, D, Lambert, G, Lee, JWook, Ferrante, T, Ma, D, Donghia, N, Fan, M, Daringer, NM, Bosch, I, Dudley, DM, O'Connor, DH, Gehrke, L, Collins, JJ|
|Date Published||2016 May 19|
|Keywords||Animals, Blood, Clustered Regularly Interspaced Short Palindromic Repeats, Computer Simulation, Dengue, Genetic Techniques, Macaca mulatta, Molecular Diagnostic Techniques, RNA, Viral, Zika Virus, Zika Virus Infection|
The 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.
|Grant List||R33 AI100190 / AI / NIAID NIH HHS / United States|