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

这笔快速反应研究(RAPID)赠款将开创一种新的诊断工具，通过减少假阴性结果，能够高精度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在高技术和低技术检测分析中实施。后者将使现场和实地检测成为可能，无需专门设备即可检测人体样本、牲畜和农业样本。正在进行的埃博拉疫情可能是历史上最大的疫情，不仅会影响西非的多个国家，还会影响到美国和其他在现场从事疾病管理和遏制工作的医护人员。虽然治疗方法和疫苗正在进行开发和测试，但尚未获得对这种威胁生命的病毒的保护或治疗。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。这项研究的成果将在全球范围内造福于经济和社会。另一个重要的好处是在一个跨学科的环境中培训和教育学者，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和公众参与度将通过已建立的“纳米人”外展计划来促进。逆转录聚合酶链式反应是一种用于在不同样本中检测埃博拉病毒的分子生物学技术。虽然这种方法具有高度灵敏的潜力，但由于处理错误，检测缺乏内部控制来否定假阴性结果。当代对照是合成的RNA转录本。这些“裸露”的RNA转录本不稳定，容易降解，因此这些分子不能可靠地用于在调查中的样本中添加；缺乏对阴性结果的核实，存在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纳米制造内在阳性对照来克服这一技术障碍，在这种对照中，合成的RNA转录物被包装成植物病毒衣壳。这种探针更接近于埃博拉病毒的RNA模板在临床或环境样本中遇到的情况。诊断探针可以在处理前添加到每个样本中，并将揭示各种样本处理步骤是否导致埃博拉病毒靶标的降解并导致假阴性读出。为应对埃博拉疫情的紧迫性，将在高科技和低技术化验中实施诊断探头，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.