Paper-based microfluidics for DNA diagnostics of malaria in low resource underserved rural communities
Paper-based microfluidics for DNA diagnostics of malaria in low resource underserved rural communities
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DOI：
10.1073/pnas.1812296116
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发表时间：
2019-03-12
影响因子：
11.1
通讯作者：
Cooper, Jonathan M.
中科院分区：
文献类型：
--
作者：
Reboud, Julien;Xu, Gaolian;Cooper, Jonathan M.
基于DNA检测的快速、低成本、物种特异性诊断在感染性疾病患者的治疗中变得越来越重要。在这里，我们展示了一项创新，即使用折纸技术实现多重、灵敏的检测，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、精确的诊断。本文提出的纸基微流体技术结合了垂直流样品处理步骤，包括全血样品制备的纸张折叠，等温扩增和侧向流检测，并结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，个人诊断在50分钟内完成（比标准的实验室PCR快）。这些测试能够从全血的手指刺血中诊断患者的疟疾种类，具有高度敏感性和特异性，在一项双盲首次人体研究中检测到98%的感染者中的疟疾。我们的方法比其他基于实地的基准技术更敏感，包括光学显微镜和行业标准快速免疫诊断测试，这两种技术都是由经验丰富的当地医疗团队进行的（分别在86%和83%的病例中检测到疟疾）。使用实时双盲参考PCR检测法对所有检测法进行独立验证。我们不仅证明了先进的、低成本的基于DNA的传感器可以在需要时在服务不足的社区实施，而且还强调了在没有实验室或基础设施的情况下开发和实施新的诊断技术所面临的挑战。
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.