A rapid and label-free platform for virus capture and identification from clinical samples
A rapid and label-free platform for virus capture and identification from clinical samples
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DOI：
10.1073/pnas.1910113117
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发表时间：
2020-01-14
影响因子：
11.1
通讯作者：
Terrones, Mauricio
中科院分区：
文献类型：
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作者：
Yeh, Yin-Ting;Gulino, Kristen;Terrones, Mauricio
新出现和重新出现的病毒是最近一些疫情暴发的原因。预测和控制疫情的一个关键步骤是及时和准确地描述新出现的病毒株。我们提出了一种便携式微流控平台，其中包含具有不同过滤孔隙率的碳纳米管阵列，用于病毒的快速浓缩和光学识别。不同的新出现的病毒株(或未知病毒)可以通过多病毒捕获组件结合表面增强拉曼光谱进行实时浓缩和鉴定。更重要的是，在芯片上捕获和检测病毒后，病毒仍然活着，并在微型设备中得到提纯，从而可以通过各种传统方法进行后续的深入表征。我们使用不同亚型的甲型禽流感病毒和呼吸道感染的人类样本验证了这一平台。这项技术成功地浓缩了鼻病毒、流感病毒和副流感病毒，并在样本中含有多种病毒时保持了化学计量病毒比例，从而模拟了混合感染。病毒捕获和检测只需几分钟，浓缩倍数增加70倍；检测只需10(2)EID50/毫升(每微升50%鸡蛋感染量)，病毒特异性为90%。使用该设备进行浓缩后，我们通过测序证明，病毒特异性读数的丰度显著增加，副流感病毒的丰度从4.1%增加到31.8%，流感病毒的丰度从0.08%增加到0.44%。这种与拉曼病毒鉴定相结合的浓缩方法构成了一种创新的系统，可以用于快速跟踪和实时监测病毒爆发。
Emerging and reemerging viruses are responsible for a number of recent epidemic outbreaks. A crucial step in predicting and controlling outbreaks is the timely and accurate characterization of emerging virus strains. We present a portable microfluidic platform containing carbon nanotube arrays with differential filtration porosity for the rapid enrichment and optical identification of viruses. Different emerging strains (or unknown viruses) can be enriched and identified in real time through a multivirus capture component in conjunction with surface-enhanced Raman spectroscopy. More importantly, after viral capture and detection on a chip, viruses remain viable and get purified in a microdevice that permits subsequent in-depth characterizations by various conventional methods. We validated this platform using different subtypes of avian influenza A viruses and human samples with respiratory infections. This technology successfully enriched rhinovirus, influenza virus, and parainfluenza viruses, and maintained the stoichiometric viral proportions when the samples contained more than one type of virus, thus emulating coinfection. Viral capture and detection took only a few minutes with a 70-fold enrichment enhancement; detection could be achieved with as little as 10(2) EID50/mL (50% egg infective dose per microliter), with a virus specificity of 90%. After enrichment using the device, we demonstrated by sequencing that the abundance of viral-specific reads significantly increased from 4.1 to 31.8% for parainfluenza and from 0.08 to 0.44% for influenza virus. This enrichment method coupled to Raman virus identification constitutes an innovative system that could be used to quickly track and monitor viral outbreaks in real time.