RAPID: Improved Detection of Ebola through Nanomanufacturing of Bio-Inspired Diagnostics

这笔快速反应研究（Rapid）赠款将率先开发一种新的诊断工具，通过减少假阴性结果，使检测致命的埃博拉病毒具有较高的准确性。通过生物工程设计和纳米制造开发的试验台将在高科技和低技术检测分析中实施。后者将能够在现场和现场进行检测，在不需要专门设备的情况下检测人类标本、牲畜和农业样品。目前的埃博拉疫情可能是历史上规模最大的一次，不仅影响到西非的多个国家，还影响到在现场进行疾病管理和控制的美国和其他医疗保健人员。虽然正在开发和测试治疗方法和疫苗，但目前还没有针对这种威胁生命的病毒的保护或治疗方法。因此，检测埃博拉病毒是预防该疾病传播的第一步，也是目前唯一的选择。这项研究的结果将在全球范围内有益于经济和社会。另一个重要的好处是在跨学科的环境中培养和教育学者，培养团队科学，连接生物工程，纳米制造，病毒学和分子生物学。K-12和公众参与将通过建立的“纳米人”外展计划来促进。逆转录聚合酶链反应是一种分子生物学技术，用于检测各种标本中的埃博拉病毒。虽然该方法具有高灵敏度的潜力，但检测缺乏内部控制来消除由于处理错误而导致的假阴性结果。当代对照是合成RNA转录物。这些“裸”RNA转录本是不稳定的，容易降解，因此这些分子不能可靠地用于刺穿正在调查的样品；缺乏对阴性结果的核查，构成了相当大的健康和环境风险。本研究旨在通过生物工程设计和纳米制造内在阳性对照来克服这一技术障碍，其中合成的RNA转录物被包装到植物病毒衣壳中。这种探针更接近于模拟埃博拉病毒RNA模板在临床或环境样本中遇到的条件。诊断探针可在处理前添加到每个标本中，并将显示各种样品处理步骤是否导致埃博拉病毒靶标降解并导致假阴性读数。为了应对埃博拉疫情的紧迫性，诊断探针将在高科技和低技术分析中实施，以便在医院以及现场和现场进行准确检测。

This Rapid Response Research (RAPID) grant will pioneer a novel diagnostic tool that will enable the detection of the deadly Ebola virus with high accuracy through reduction of false negative results. The test-bed developed through bioengineering design and nanomanufacturing will be implemented in high-tech and low-tech detection assays. The latter will enable detection on-site and in-field, testing human specimens, livestock and agricultural samples without the need for specialized equipment. The ongoing Ebola epidemic could potentially be the largest in history, not only affecting multiple countries in West Africa, but also U.S. and other healthcare personnel working on-site in disease management and containment. While therapeutics and vaccines are undergoing development and testing, protection or treatment of this life-threatening virus is not yet available. Therefore, detection of Ebola virus is the first step, and currently the only option, to prevent the spread of the disease. Results from this research will benefit the economy and society on a global scale. An important further benefit is the training and education of scholars in an interdisciplinary environment fostering team sciences, bridging bioengineering, nanomanufacturing, virology, and molecular biology. K-12 and public engagement will be promoted through the established 'The Nanoman' outreach program. Reverse transcription polymerase chain reaction is a molecular biological technique used to detect Ebola virus in diverse specimens. While this method has the potential to be highly sensitive, the assays lack internal controls to negate false negative results due to processing errors. Contemporary controls are synthetic RNA transcripts. These 'naked' RNA transcripts are unstable and prone to degradation, therefore these molecules cannot be reliably used to spike samples under investigation; the lack of verification of negative results presents a considerable health and environmental risk. This research sets out to overcome this technological hurdle through bioengineering design and nanomanufacturing of an intrinsic positive control, in which the synthetic RNA transcripts are packaged into a plant virus capsid. Such a probe mimics more closely the conditions encountered by the RNA template of the Ebola virus within clinical or environmental samples. The diagnostic probe can be added to each specimen prior to processing and will reveal whether various sample-processing steps have resulted in degradation of the Ebola virus target and resulted in a false negative read-out. In response to the urgency of the Ebola epidemic, the diagnostic probe will be implemented in high-tech and low-tech assays to enable accurate detection in hospitals and in-field and on-site.