Development and Commercialization of Analyte Specific Reagents (ASRs) for the Diagnosis of Selected Arthropod-Borne Viruses on FDA-Cleared Real-time P

Phase I will develop multiple combinations of qRT-PCR primers and probes for some of six diseases of military importance: Dengue, Rift Valley Fever, Chikungunya, Crimean-Congo Hemorrhagic Fever, Sandfly Fever, and Tick-Borne Encephalitis. Based on known sequence information from the literature and contacts at disease labs, probes and primer pairs will be designed and synthesized for testing; expected probe formats include hydrolysis probes, scorpions, and molecular beacons. Positive control plasmids containing target amplicon sequences will be designed, synthesized, and sequence-verified. Assays will be optimized for cycle profile, magnesium concentration, presence of competing nucleic acids, melting temperature, and gels to verify predicted amplicon length. Specificity and sensitivity will be established. Final probes and primer pairs will be provided to a Army Testing Lab for evaluation against disease targets. Acceptable probes and primers will be manufactured under cGMP, and a minimum of 100 vials of at least one analyte assay provided to the military for testing; if successful, the expectation is assays for three of the six diseases by the end of Phase I Option.