Combined Viral Load and Ultrasensitive Serology Panel for Rapid Ebola Diagnostics

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 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.