Combined Viral Load and Ultrasensitive Serology Panel for Rapid Ebola Diagnostics

 描述（由申请方提供）：诊断病毒性出血热（VHF）病原体，尤其是埃博拉病毒、马尔堡病毒和拉沙病毒，对人类和非人灵长类动物均具有高度致死性。埃博拉病毒和马尔堡病毒在世界各地，特别是卫生基础设施薄弱的地区造成蔓延性疫情。目前，有记录以来最大的埃博拉疫情正在西非爆发，但目前疫情的初步诊断发生在欧洲。这使人们认识到，需要新的、更简单的诊断方法。一种新的生物传感平台，称为SP-IRIS，允许对结合到用特异性捕获抗体功能化的传感器表面的单个病毒颗粒进行无标记计数，将提供灵敏度检测埃博拉病毒的能力，而不需要显著的医疗基础设施。SP-IRIS已经显示出使用在人全血中掺入的VSV-假型病毒进行埃博拉病毒检测的高灵敏度和特异性。在这项拟议的工作中，将使用更好地模拟真实埃博拉病毒颗粒形状的埃博拉病毒样颗粒进一步开发埃博拉检测试剂，以证明使用安全和相似模型的开发。SP-IRIS传感器还可以用作显微镜系统，可以使结合在传感器上的埃博拉病毒颗粒可视化。因此，第一个目标是开发一种自动化软件分析算法，其将能够识别结合到传感器表面的丝状埃博拉颗粒，作为提供正交度量以提高测定的灵敏度和特异性的方式。该算法将在真正的埃博拉病毒上进行验证。在第二个目标中，将研究在SP-IRIS芯片上添加埃博拉抗原。成功纳入这种超灵敏抗体检测将使埃博拉病毒载量和血清学检测相结合。综合测试将提供关于潜在感染者如何应对疾病的重要信息。最后，将评价检测试剂盒的稳定性和有效期。所提出的工作的成功将证明SP-IRIS可以在需要时克服传统诊断技术的障碍。我们设想，SP-IRIS在未来可以作为诊断平台，可以运行多重病毒性出血热面板，并结合血清学直接从一个小的血液样本在一个样本到答案的格式在低成本，它将是发展中国家和国家参考实验室也有用。成功的结果将为第二阶段的建议注入信心，建立一个全面的病毒性出血热小组，包括更常见的出血热，即拉沙热，马尔堡，登革热和黄热病以及具有类似症状的流行病，即疟疾和基孔肯雅热。

 DESCRIPTION (provided by applicant): Diagnostics Viral Hemorrhagic Fever (VHF) pathogens, especially Ebola, Marburg, and Lassa, are highly lethal for both humans and nonhuman primates. Ebola and Marburg cause propagated outbreaks in areas of the world, especially those with poor health infrastructures. Currently, the largest outbreak of Ebola ever recorded is ongoing in West Africa but the initial diagnosis of the current outbreak occurred in Europe. This drives home the fact that new, simpler diagnostic approaches are needed. A novel biosensing platform, termed SP-IRIS that allows label-free counting of individual virus particles bound to the sensor surface functionalized with specific capture antibodies would provide the ability to sensitivity detect the Ebola virus without needing a significant healthcare infrastructue. SP-IRIS has shown high sensitivity and specificity for Ebola detection using VSV- pseudotype virus spiked in human whole blood. In this proposed work, further development of the Ebola assay, using Ebola viral-like particles that will better mimic bone fide Ebola virus particle shape will be used to demonstrate development using a safe and similar model. The SP-IRIS sensor can also serve as a microscopy system that can visualize the Ebola viral particle bound on the sensor. Thus, the first aim is to develop an automated software analysis algorithm that will be able to identify filamentous Ebola particles bound to the sensor surface as a way to provide an orthogonal metric to improve sensitivity and specificity of the assay. This algorithm will then be verified on bone fide Ebola virus. In the second aim the addition of Ebola antigen on the SP-IRIS chip will be studied. Successful inclusion of this ultrasensitive antibody test will enable a combined Ebola viral load and serology test. The combined test will provide important information on how a potentially infected individual is responding to the disease. Finally, the stability and shelf life of the assay will be evaluated. The success of the proposed work will prove that SP-IRIS can overcome the hurdles of traditional diagnostic technologies at the point-of-need. We envision that SP-IRIS in the future can serve as diagnostic platform that can run a multiplexed viral hemorrhagic fever panel and combined with serology directly from a small blood sample in a sample-to-answer format at a low-cost that it will be accessible to developing countries and also useful for national reference labs. Successful results will instill confidence towards a Phase II proposal to create a comprehensive viral hemorrhagic fever panel that will include more common hemorrhagic fevers, i.e. Lassa, Marburg, Dengue, and Yellow Fever as well as prevalent diseases with similar symptoms, i.e. Malaria and Chikungunya.