Division of Biomedical Engineering, University of Glasgow, G12 8LT Glasgow, United Kingdom.
Nano Biomedical Research Centre, School of Biomedical Engineering, Shanghai Jiao Tong University, 200030 Shanghai, People's Republic of China.
Proc Natl Acad Sci U S A. 2019 Mar 12;116(11):4834-4842. doi: 10.1073/pnas.1812296116. Epub 2019 Feb 19.
Rapid, low-cost, species-specific diagnosis, based upon DNA testing, is becoming important in the treatment of patients with infectious diseases. Here, we demonstrate an innovation that uses origami to enable multiplexed, sensitive assays that rival polymerase chain reactions (PCR) laboratory assays and provide high-quality, fast precision diagnostics for malaria. The paper-based microfluidic technology proposed here combines vertical flow sample-processing steps, including paper folding for whole-blood sample preparation, with an isothermal amplification and a lateral flow detection, incorporating a simple visualization system. Studies were performed in village schools in Uganda with individual diagnoses being completed in <50 min (faster than the standard laboratory-based PCR). The tests, which enabled the diagnosis of malaria species in patients from a finger prick of whole blood, were both highly sensitive and specific, detecting malaria in 98% of infected individuals in a double-blind first-in-human study. Our method was more sensitive than other field-based, benchmark techniques, including optical microscopy and industry standard rapid immunodiagnostic tests, both performed by experienced local healthcare teams (which detected malaria in 86% and 83% of cases, respectively). All assays were independently validated using a real-time double-blinded reference PCR assay. We not only demonstrate that advanced, low-cost DNA-based sensors can be implemented in underserved communities at the point of need but also highlight the challenges associated with developing and implementing new diagnostic technologies in the field, without access to laboratories or infrastructure.
基于 DNA 检测的快速、低成本、物种特异性诊断在传染病患者的治疗中变得越来越重要。在这里，我们展示了一种创新，它使用折纸技术实现了多重、敏感的检测，可与聚合酶链反应（PCR）实验室检测相媲美，并为疟疾提供高质量、快速的精准诊断。这里提出的基于纸张的微流控技术结合了垂直流样品处理步骤，包括用于全血样品制备的折纸，以及等温扩增和横向流动检测，同时还整合了一个简单的可视化系统。研究在乌干达的乡村学校进行，每个诊断的完成时间不到 50 分钟（比基于实验室的标准 PCR 更快）。这些测试能够从指尖刺取的全血中诊断出患者的疟疾种类，具有高度的敏感性和特异性，在一项双盲首次人体研究中，能够检测到 98%的感染个体中的疟疾。我们的方法比其他基于现场的基准技术更敏感，包括光学显微镜和行业标准的快速免疫诊断测试，这两种方法都是由经验丰富的当地医疗团队进行的（分别检测到 86%和 83%的病例中的疟疾）。所有的检测都是独立使用实时双盲参考 PCR 检测进行验证的。我们不仅证明了先进的、低成本的基于 DNA 的传感器可以在没有实验室或基础设施的情况下在需要的地方在服务不足的社区中实施，还强调了在没有实验室或基础设施的情况下，在现场开发和实施新诊断技术所面临的挑战。
