MMN laboratory CNRS UMR7083 Gulliver, ESPCI Paris, PSL Research University, Paris, France.
Institut Pasteur, HIV, Inflammation and Persistence Unit, Paris, France.
Sci Rep. 2017 May 2;7(1):1347. doi: 10.1038/s41598-017-00758-9.
The most performing techniques enabling early diagnosis of infectious diseases rely on nucleic acid detection. Today, because of their high technicality and cost, nucleic acid amplification tests (NAAT) are of benefit only to a small fraction of developing countries population. By reducing costs, simplifying procedures and enabling multiplexing, paper microfluidics has the potential to considerably facilitate their accessibility. However, most of the studies performed in this area have not quit the lab. This letter brings NAAT on paper closer to the field, by using clinical samples and operating in a resource-limited setting. We first performed isothermal reverse transcription and Recombinase Polymerase Amplification (RT-RPA) of synthetic Ribonucleic Acid (RNA) of Ebola virus using paper microfluidics devices. We further applied this method in Guinea to detect the presence of Ebola virus in human sample RNA extracts, with minimal facilities (carry-on detection device and freeze-dried reagents on paper). RT-RPA results were available in few minutes and demonstrate a sensitivity of 90.0% compared to the gold-standard RT-PCR on a set of 43 patient samples. Furthermore, the realization of a nine-spot multilayered device achieving the parallel detection of three distinct RNA sequences opens a route toward the detection of multiple viral strains or pathogens.
最常用于传染病早期诊断的技术依赖于核酸检测。如今，由于其技术要求高、成本高，核酸扩增检测（NAAT）仅对一小部分发展中国家的人口有益。通过降低成本、简化程序并实现多重检测，纸基微流控技术有可能极大地促进其普及。然而，该领域的大多数研究仍停留在实验室阶段。这封信通过使用临床样本并在资源有限的环境中运行，使基于纸的 NAAT 更接近现场。我们首先使用纸基微流控设备对埃博拉病毒的合成核糖核酸（RNA）进行等温逆转录和重组酶聚合酶扩增（RT-RPA）。我们进一步在几内亚应用该方法检测人类样本 RNA 提取物中埃博拉病毒的存在，仅使用基本设施（手提式检测设备和冻干试剂在纸上）。几分钟内即可获得 RT-RPA 结果，与 43 个患者样本的金标准 RT-PCR 相比，其灵敏度为 90.0%。此外，实现一种具有九个斑点的多层器件，可平行检测三种不同的 RNA 序列，为同时检测多种病毒株或病原体开辟了道路。
