Rapid, Low-Cost Detection of Zika Virus Using Programmable Biomolecular Components
Rapid, Low-Cost Detection of Zika Virus Using Programmable Biomolecular Components
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DOI：
10.1016/j.cell.2016.04.059
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发表时间：
2016-05-19
期刊：
影响因子：
64.5
通讯作者：
Collins, James J.
中科院分区：
文献类型：
--
作者：
Pardee, Keith;Green, Alexander A.;Collins, James J.
最近的寨卡病毒暴发突出表明，需要能够迅速开发的低成本诊断方法，以便在大流行地区分发和使用。在这里，我们报告了一种用于检测寨卡病毒RNA基因组的无细胞、纸质传感器的快速设计、组装和验证的管道。通过将等温RNA扩增与脚点开关RNA传感器连接起来，我们检测了临床相关的寨卡病毒序列浓度，并证明了对密切相关的登革热病毒序列的特异性。当与新的基于CRISPR/ cas9的模块相结合时，我们的传感器可以以单碱基分辨率区分病毒株。我们成功地演示了一种简单的现场样品处理工作流程，并从病毒血症猕猴的血浆中检测出寨卡病毒。我们的冻干生物分子平台解决了在该领域部署分子诊断的重要实际限制，并展示了如何使用合成生物学来开发应对全球健康危机的诊断工具。
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.