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ARS Home » Research » Research Project #431003

Research Project: Molecular Analytical Methods Development to Support Arbovirus Research

Location: Foreign Arthropod Borne Animal Disease Research

Project Number: 3022-32000-022-023-S
Project Type: Non-Assistance Cooperative Agreement

Start Date: Sep 1, 2016
End Date: Aug 31, 2021

Objective 1: Immunochemistry of Virus neutralization. In all of the NP103 Animal Health Projects, there will be a requirement to optimize antibody mediated virus neutralization assays, develop methods to identify primary/linear and secondary/conformational epitopes of infectious viruses, and evaluate the immunogenicity of identified epitope structures. Objective 2: Generate Reverse Genetic derived recombinant viruses. In the NP103 Animal Health Project related to bunyaviruses there is a goal to identify viral molecular determinants of virulence and mechanisms of viral pathogenesis, which will require the generation of multiple recombinant viruses. Objective 3: Development of vaccines and differentiating infected from vaccinated animal (DIVA) compatible diagnostics. In all the NP103 Animal Heath Projects, there is an end goal of detection and control of endemic and potentially emerging arboviruses, which will require new diagnostic tools and vaccines.

Objective 1: A. Establish assay standards, controls and methods for antibody-mediated virus neutralization for selected Flavivirus and Orbivirus agents. This work will include supportive immunological assays as needed. Reagents will come from existing stocks of viruses, sera and other materials as well as from animal studies to be completed as parts of other studies and through multiple collaborations. B. Evaluate potential neutralization determinants of the selected viruses that may be generated via computational platforms, genetic analysis, immunoprecipitation or other methods. The specific goals will be to identify primary/linear and secondary/conformational epitopes associated with antibody-mediated neutralization. Objective 2: A. Establish the standard operating procedures for generation of recombinant Rift Valley fever viruses. This work will include supportive virological assays and sequencing to generate a titered working stock and confirmation. B. Assist in evaluation of selected recombinant viruses generated in small and target animal species. The specific goal is to identify specific genetic markers associated with virulence to better predict outcome of an outbreak and improve vaccine development. Objective 3: A. Develop new arbovirus control strategies using modern vaccine technology that allows differentiating infected from vaccinated animals (DIVA). B. Develop the reagent and assays compatible with DIVA control strategy.