Paper-based RNA detection and multiplexed analysis for Ebola virus diagnostics.
Paper-based RNA detection and multiplexed analysis for Ebola virus diagnostics.
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DOI：
10.1038/s41598-017-00758-9
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发表时间：
2017-05-02
影响因子：
4.6
通讯作者：
Tabeling P
中科院分区：
文献类型：
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作者：
Magro L;Jacquelin B;Escadafal C;Garneret P;Kwasiborski A;Manuguerra JC;Monti F;Sakuntabhai A;Vanhomwegen J;Lafaye P;Tabeling P
能够早期诊断传染病的最有效的技术依赖于核酸检测。今天，由于其高技术性和高成本，核酸扩增测试(NAAT)只对一小部分发展中国家人口有利。通过降低成本、简化程序和实现多路复用，纸张微流体有可能极大地便利它们的可及性。然而，在这一领域进行的大多数研究并没有退出实验室。这封信通过使用临床样本并在资源有限的环境下操作，使纸面上的NAAT更接近现场。我们首次利用纸质微流控装置对合成的埃博拉病毒核糖核酸(RNA)进行了等温逆转录和重组酶聚合酶扩增(RT-RPA)。我们进一步将这种方法应用于几内亚，以最小的设施(随身检测设备和纸上的冻干试剂)检测人类样本RNA提取液中是否存在埃博拉病毒。RT-RPA结果在几分钟内就可以得到，在43例患者样本中，与金标准RT-PCR相比，RT-RPA结果的敏感性为90.0%。此外，实现对三个不同RNA序列的并行检测的九点多层设备的实现，为检测多个病毒株或病原体开辟了一条途径。
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.