title: Rapid, Low-Cost Detection of Zika Virus Using Programmable Biomolecular Components.
abstract: 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.
authors.affiliation: Leslie Dan Faculty of Pharmacy, University of Toronto, Toronto, ON M5S 3M2, Canada.
authors.forename: Keith
authors.initials: K
authors.lastname: Pardee
authors.affiliation: Biodesign Center for Molecular Design and Biomimetics, The Biodesign Institute and the School of Molecular Sciences, Arizona State University, AZ 85287, USA.
authors.forename: Alexander A
authors.initials: AA
authors.lastname: Green
authors.affiliation: Institute for Medical Engineering & Science, Massachusetts Institute of Technology, Cambridge, MA 02139, USA.
authors.forename: Melissa K
authors.initials: MK
authors.lastname: Takahashi
authors.affiliation: Institute for Medical Engineering & Science, Massachusetts Institute of Technology, Cambridge, MA 02139, USA; Department of Biomedical Engineering, Boston University, Boston, MA 02215, USA; Wyss Institute for Biologically Inspired Engineering, Harvard University, Boston, MA 02115, USA.
authors.forename: Dana
authors.initials: D
authors.lastname: Braff
authors.affiliation: Wyss Institute for Biologically Inspired Engineering, Harvard University, Boston, MA 02115, USA; School of Applied and Engineering Physics, Cornell University, Ithaca, NY 14853, USA.
authors.forename: Guillaume
authors.initials: G
authors.lastname: Lambert
authors.affiliation: Wyss Institute for Biologically Inspired Engineering, Harvard University, Boston, MA 02115, USA.
authors.forename: Jeong Wook
authors.initials: JW
authors.lastname: Lee
authors.affiliation: Wyss Institute for Biologically Inspired Engineering, Harvard University, Boston, MA 02115, USA.
authors.forename: Tom
authors.initials: T
authors.lastname: Ferrante
authors.affiliation: Biodesign Center for Molecular Design and Biomimetics, The Biodesign Institute and the School of Molecular Sciences, Arizona State University, AZ 85287, USA.
authors.forename: Duo
authors.initials: D
authors.lastname: Ma
authors.affiliation: Wyss Institute for Biologically Inspired Engineering, Harvard University, Boston, MA 02115, USA.
authors.forename: Nina
authors.initials: N
authors.lastname: Donghia
authors.affiliation: Addgene, Cambridge, MA 02139, USA.
authors.forename: Melina
authors.initials: M
authors.lastname: Fan
authors.affiliation: Institute for Medical Engineering & Science, Massachusetts Institute of Technology, Cambridge, MA 02139, USA.
authors.forename: Nichole M
authors.initials: NM
authors.lastname: Daringer
authors.affiliation: Institute for Medical Engineering & Science, Massachusetts Institute of Technology, Cambridge, MA 02139, USA.
authors.forename: Irene
authors.initials: I
authors.lastname: Bosch
authors.affiliation: Wisconsin National Primate Research Center and Department of Pathology and Laboratory Medicine, UW-Madison, Madison, WI 53706, USA.
authors.forename: Dawn M
authors.initials: DM
authors.lastname: Dudley
authors.affiliation: Wisconsin National Primate Research Center and Department of Pathology and Laboratory Medicine, UW-Madison, Madison, WI 53706, USA.
authors.forename: David H
authors.initials: DH
authors.lastname: O'Connor
authors.affiliation: Institute for Medical Engineering & Science, Massachusetts Institute of Technology, Cambridge, MA 02139, USA; Department of Microbiology and Immunobiology, Harvard Medical School, Boston, MA 02115, USA; Harvard-MIT Program in Health Sciences and Technology, Cambridge, MA 02139, USA.
authors.forename: Lee
authors.initials: L
authors.lastname: Gehrke
authors.affiliation: Institute for Medical Engineering & Science, Massachusetts Institute of Technology, Cambridge, MA 02139, USA; Wyss Institute for Biologically Inspired Engineering, Harvard University, Boston, MA 02115, USA; Harvard-MIT Program in Health Sciences and Technology, Cambridge, MA 02139, USA; Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, MA 02139, USA; Synthetic Biology Center, Massachusetts Institute of Technology, Cambridge, MA 02139, USA; Broad Institute of MIT and Harvard, Cambridge, MA 02142, USA. Electronic address: jimjc@mit.edu.
authors.forename: James J
authors.initials: JJ
authors.lastname: Collins
mesh_terms: D000818:Animals; D001769:Blood / Q000821:virology; D064112:Clustered Regularly Interspaced Short Palindromic Repeats; D003198:Computer Simulation; D003715:Dengue / Q000175:diagnosis / Q000821:virology; D005821:Genetic Techniques; D008253:Macaca mulatta; D025202:Molecular Diagnostic Techniques / Q000191:economics / Q000379:methods*; D012367:RNA, Viral / Q000302:isolation & purification; D000071244:Zika Virus / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D000071243:Zika Virus Infection / Q000175:diagnosis* / Q000821:virology
chemical_list: D012367:RNA, Viral