TY - JOUR
T1 - Rapid, Low-Cost Detection of Zika Virus Using Programmable Biomolecular Components
AU - Pardee, Keith
AU - Green, Alexander
AU - Takahashi, Melissa K.
AU - Braff, Dana
AU - Lambert, Guillaume
AU - Lee, Jeong Wook
AU - Ferrante, Tom
AU - Ma, Duo
AU - Donghia, Nina
AU - Fan, Melina
AU - Daringer, Nichole M.
AU - Bosch, Irene
AU - Dudley, Dawn M.
AU - O'Connor, David H.
AU - Gehrke, Lee
AU - Collins, James J.
N1 - Funding Information:
We would like to thank Marcelle Tuttle from the Church Lab (Wyss Institute) for the vectors used to produce the lentivirus. We would also like to thank Xiao Tan and Shimyn Slomovic for helpful comments on the manuscript, as well as Ewen Cameron, Andres Cubillos, James Niemi and Dionna Williams for assistance with project logistics. The work was supported by the Wyss Institute for Biologically Inspired Engineering, MIT’s Center for Microbiome Informatics and Therapeutics, and the Defense Threat Reduction Agency grant HDTRA1-14-1-0006. A.A.G acknowledges startup funds provided by Arizona State University. L.G. acknowledges support from NIH AI100190.
Publisher Copyright:
© 2016 Elsevier Inc.
PY - 2016/5/19
Y1 - 2016/5/19
N2 - Summary 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.
AB - Summary 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.
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U2 - 10.1016/j.cell.2016.04.059
DO - 10.1016/j.cell.2016.04.059
M3 - Article
C2 - 27160350
AN - SCOPUS:84973486576
VL - 165
SP - 1255
EP - 1266
JO - Cell
JF - Cell
SN - 0092-8674
IS - 5
ER -