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

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Final rept. 1 Jan 2011-31 Aug 2012

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The primary goal of the Phase II Contract was to continue to design, manufacture, and test real time polymerase chain reaction qPCR reagents for detection of one or more of six potentially deadly arbovirus pathogens Dengue, Rift Valley Fever, Chikungunya, Crimean-Congo Hemorrhagic Fever, Sand Fly Fever, and Tick-Borne Encephalitis viruses. The qPCR reagents being provided in Phase II are additional primers and probes signature sets sets for qPCR applications, as well as positive controls synthetic targets for the assays. Work product in Phase II included 1 design and synthesis of a synthetic target representing all four Dengue serotypes 2 cloning of a plasmid control for the new RT-qPCR signature set 4 qPCR testing of the new Dengue signature set, synthetic template, and plasmid control 5 determination of efficiency, Linearity of Range LOR, and Limit of Detection LOD for all eight qPCR signature sets 6 bioinformatic analysis and alignment of sequences published for the high priority agents listed 7 design and synthesis of a new consensus qPCR signature set for Dengue types 1-4 DENcon based on bioinformatics analysis of 42 Dengue subtypes 8 design and synthesis of a synthetic template for the consensus Dengue signature set 9 design and synthesis of a control plasmid ARBO9 containing sites for all eight signature sets 10 the testing of RNA generated from ARBO9 as a synthetic target and 11 stability testing of all reagents, including positive controls.

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  • Medicine and Medical Research

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