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

这项快速反应研究（RAPID）拨款将开创一种新的诊断工具，通过减少假阴性结果，能够高准确性地检测致命的埃博拉病毒。通过生物工程设计和纳米制造开发的试验台将用于高技术和低技术检测分析。后者将能够在现场和实地进行检测，测试人体标本、牲畜和农业样本，而不需要专门设备。正在进行的埃博拉疫情可能是历史上最大的一次，不仅影响西非的多个国家，而且还影响到在现场进行疾病管理和遏制的美国和其他医护人员。虽然治疗方法和疫苗正在开发和测试中，但这种威胁生命的病毒的保护或治疗尚不可用。因此，检测埃博拉病毒是防止疾病传播的第一步，也是目前唯一的选择。这项研究的成果将有利于全球范围内的经济和社会。另一个重要的好处是在跨学科环境中培养和教育学者，培养团队科学，桥接生物工程，纳米制造，病毒学和分子生物学。K-12和公众参与将通过既定的“纳米人”外展计划来促进。逆转录聚合酶链反应是一种用于检测不同标本中埃博拉病毒的分子生物学技术。虽然该方法具有高灵敏度的潜力，但该检测试剂盒缺乏内部对照，无法消除由于处理错误导致的假阴性结果。当代对照是合成RNA转录物。这些“裸”RNA转录物不稳定且易于降解，因此这些分子不能可靠地用于加标研究中的样品;缺乏对阴性结果的验证会带来相当大的健康和环境风险。这项研究旨在通过生物工程设计和内在阳性对照的纳米制造来克服这一技术障碍，其中合成的RNA转录本被包装到植物病毒衣壳中。这种探针更接近地模拟了临床或环境样本中埃博拉病毒RNA模板所遇到的条件。诊断探针可以在处理之前添加到每个标本中，并将揭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，诊断探针将在高技术和低技术含量的检测中实施，以实现医院和现场的准确检测。

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.