Combined Viral Load and Ultrasensitive Serology Panel for Rapid Ebola Diagnostics

用于快速埃博拉诊断的病毒载量和超灵敏血清学组合组合

• 批准号: 8906221
• 负责人: David Scott Freedman
• 金额: $ 22.41万
• 依托单位: NEXGEN ARRAYS, LLC
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-03-01 至 2016-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8906221
• 关键词: Acute; Africa; Algorithms; Animal Model; Animals; Antibodies; Antigens; Area; Automation; Binding; Biological Assay; Biological Response Modifier Therapy; Biosensing Techniques; Blood specimen; Boston; Cessation of life; Chronic; Complex; Computer software; Dengue; Detection; Developing Countries; Development; Diagnosis; Diagnostic; Disease; Disease Management; Disease Outbreaks; Ebola virus; Enzyme-Linked Immunosorbent Assay; Europe; Event; Filovirus; Frankfurt-Marburg Syndrome Virus; Future; Goals; Health; Healthcare; Home environment; Human; Image; Immunoassay; Immunoglobulin G; Immunoglobulin M; Individual; Infection; Label; Life; Malaria; Measures; Mentors; Methods; Microscopy; Modality; Modeling; Pathogen detection; Patient Care; Patients; Phase; Probability; Procedures; Production; Proteins; Research Infrastructure; Running; Sampling; Sensitivity and Specificity; Serologic tests; Serological; Shapes; Solid; Specificity; Spottings; Surface; Symptoms; System; Technology; Test Result; Testing; Time; Training; Universities; Vesicular stomatitis Indiana virus; Viral; Viral Antibodies; Viral Hemorrhagic Fevers; Viral Load result; Virion; Virus; Virus Diseases; Whole Blood; Work; Yellow Fever; assay development; base; bone; chikungunya; clinically relevant; cost; design; follow-up; graduate student; improved; instrument; instrumentation; nanoparticle; new technology; nonhuman primate; novel; particle; pathogen; public health relevance; rapid detection; research study; response; screening; sensor; software development; spectrograph; success; viral detection

 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.

 描述（由申请人提供）：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，对人类和非人类灵长类动物具有高度致命性。埃博拉病毒和马尔堡病毒在世界各地引起疫情蔓延，特别是那些卫生基础设施薄弱的地区。目前，有记录以来最大规模的埃博拉疫情正在西非爆发，但当前疫情的初步诊断发生在欧洲。这说明我们需要新的、更简单的诊断方法。一种名为 SP-IRIS 的新型生物传感平台，可以对与特定捕获抗体功能化的传感器表面结合的单个病毒颗粒进行无标记计数，从而无需大量医疗基础设施即可灵敏地检测埃博拉病毒。 SP-IRIS 使用掺入人全血中的 VSV 假型病毒检测埃博拉病毒显示出高灵敏度和特异性。在这项拟议的工作中，进一步开发埃博拉检测，使用埃博拉病毒样颗粒，更好地模拟真实的埃博拉病毒颗粒形状，将用于证明使用安全且相似的模型进行开发。 SP-IRIS 传感器还可以用作显微镜系统，可以可视化传感器上结合的埃博拉病毒颗粒。因此，第一个目标是开发一种自动化软件分析算法，能够识别与传感器表面结合的丝状埃博拉病毒颗粒，作为提供正交度量的一种方式，以提高测定的灵敏度和特异性。然后，该算法将在真正的埃博拉病毒上进行验证。第二个目标将研究在 SP-IRIS 芯片上添加埃博拉抗原。成功纳入这种超灵敏抗体测试将使埃博拉病毒载量和血清学测试相结合。联合测试将提供有关潜在感染者如何应对该疾病的重要信息。最后，将评估测定的稳定性和保质期。这项工作的成功将证明 SP-IRIS 可以根据需要克服传统诊断技术的障碍。我们预计，未来 SP-IRIS 可以作为诊断平台，运行多重病毒性出血热检测，并以低成本的方式直接从少量血液样本中以样本到答案的形式与血清学相结合，以便发展中国家可以使用，也对国家参考实验室有用。成功的结果将为第二阶段提案注入信心，该提案旨在创建一个全面的病毒性出血热小组，其中包括更常见的出血热，即拉沙热、马尔堡病毒、登革热和黄热病，以及具有类似症状的流行疾病，即疟疾和基孔肯雅热。