Infectious disease outbreaks such as Ebola and other Viral Hemorrhagic Fevers (VHF) require low-complexity, specific, and differentiated diagnostics as illustrated by the recent outbreak in the Democratic Republic of Congo. Here, we describe amplification-free spectrally multiplex detection of four different VHF total RNA samples using multi-spot excitation on a multimode interference waveguide platform along with combinatorial fluorescence labeling of target nucleic acids. In these experiments, we observed an average of 8-fold greater fluorescence signal amplitudes for the Ebola total RNA sample compared to three other total RNA samples: Lake Victoria Marburg Virus, Ravn Marburg Virus, and Crimean-Congo Hemorrhagic Fever. We have attributed this amplitude amplification to an increased amount of RNA during synthesis of soluble glycoprotein in infection. This hypothesis is confirmed by single molecule detection of the total RNA sample after heat-activated release from the carrier microbeads. From these experiments, we observed at least a 5.3x higher RNA mass loading on the Ebola carrier microbeads compared to the Lake Victoria Marburg carrier microbeads, which is consistent with the known production of soluble glycoprotein during infection.

中文翻译：

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病毒性出血热的光流控无放大多重检测

埃博拉病毒和其他病毒性出血热 (VHF) 等传染病暴发需要低复杂性、特异性和差异化的诊断，如刚果民主共和国最近的暴发所示。在这里，我们描述了使用多模干涉波导平台上的多点激发以及靶核酸的组合荧光标记对四种不同的 VHF 总 RNA 样品进行无扩增光谱多重检测。在这些实验中，我们观察到埃博拉病毒总 RNA 样本的荧光信号幅度平均比其他三个总 RNA 样本高 8 倍：维多利亚湖马尔堡病毒、拉文马尔堡病毒和克里米亚-刚果出血热。我们将这种幅度放大归因于感染中可溶性糖蛋白合成过程中 RNA 量的增加。这一假设通过从载体微珠热激活释放后对总 RNA 样品的单分子检测得到证实。从这些实验中，我们观察到与维多利亚湖马尔堡载体微珠相比，埃博拉病毒载体微珠上的 RNA 负载量至少高出 5.3 倍，这与已知的感染期间可溶性糖蛋白的产生一致。