Biomedical Institute for Global Health Research and Technology, National University of Singapore, Singapore, 117599, Singapore.
Institute of Molecular and Cell Biology, Agency for Science, Technology and Research, Singapore, 138673, Singapore.
Nat Commun. 2018 Aug 13;9(1):3238. doi: 10.1038/s41467-018-05733-0.
Rapid, visual detection of pathogen nucleic acids has broad applications in infection management. Here we present a modular detection platform, termed enzyme-assisted nanocomplexes for visual identification of nucleic acids (enVision). The system consists of an integrated circuit of enzyme-DNA nanostructures, which function as independent recognition and signaling elements, for direct and versatile detection of pathogen nucleic acids from infected cells. The built-in enzymatic cascades produce a rapid color readout for the naked eye; the assay is thus fast (<2 h), sensitive (<10 amol), and readily quantified with smartphones. When implemented on a configurable microfluidic platform, the technology demonstrates superior programmability to perform versatile computations, for detecting diverse pathogen targets and their virus-host genome integration loci. We further design the enVision platform for molecular-typing of infections in patient endocervical samples. The technology not only improves the clinical inter-subtype differentiation, but also expands the intra-subtype coverage to identify previously undetectable infections.
快速、可视化的病原体核酸检测在感染管理中有广泛的应用。在这里,我们提出了一种模块化的检测平台,称为酶辅助纳米复合物用于核酸的可视化识别(enVision)。该系统由酶-DNA 纳米结构的集成电路组成,作为独立的识别和信号元件,可直接、多样地检测感染细胞中的病原体核酸。内置的酶级联反应产生肉眼可见的快速颜色读数;因此,该检测方法快速(<2 小时)、灵敏(<10 飞摩尔),并且可以用智能手机进行轻松定量。当在可配置的微流控平台上实现时,该技术具有卓越的可编程性,可以执行各种病原体靶标及其病毒-宿主基因组整合位点的多样化计算。我们进一步将 enVision 平台设计用于患者宫颈内样本的感染分子分型。该技术不仅提高了临床亚型间的区分度,还扩大了亚型内的覆盖范围,以识别以前无法检测到的感染。