RAPID: Highly Specific and Sensitive Detection of Ebola Virus from Blood

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1509759 - SantangeloIn order to mitigate the spread of Ebola infections, detecting the virus in patients before the onset of symptoms is imperative. This requires, fast, sensitive and specific detection of the virus. Current methods lack sensitivity and ease of use. This project will adapt the author's previous method for detecting ultra low Ebola concentration in patient's blood. Two new methods will be developed; one can be used in the field and the second one, which is more rigorous, is suitable for laboratory tests. These methods are expected to reduce the rate at which Ebola epidemic spreads.Currently quantitative PCR is the state-of-the-art for Ebola detection, typically from blood samples. These tests though, are time intensive and require a high degree of expertise to perform. Further the method is useful when viral concentration is relatively high in the patient. Given the need to test large numbers of people as a part of quarantine strategies, improved early diagnostics are imperative. Therefore the aim of the project is to develop a more sensitive approach to detecting Ebola viral RNA (vRNA) and single viral particles. The research team will develop a single tube assay for detecting viral RNA in whole blood, suitable for field application. In addition, a slide-based assay for detecting single viral particles will also be developed, through the detection of vRNA-protein interactions, with single virion sensitivity suitable for a laboratory setting. In both cases, peptide labeled, single RNA sensitive, RNA imaging probes and proximity ligation assay (PLA) methods will be utilized. The results will be compared with current methods to demonstrate the improved sensitivity. The combinations of both site-specific probe binding and distance-dependent detection and amplification, will ensure both specificity (zero errors) and sensitivity (10 copies/ml).