Location: Foreign Arthropod Borne Animal Disease Research
Project Number: 3022-32000-024-022-S
Project Type: Non-Assistance Cooperative Agreement
Start Date: Sep 1, 2026
End Date: Aug 31, 2028
Objective:
The overarching objective of this agreement is to develop, optimize and validate high-specificity competitive ELISA antibody test for detection and surveillance of transboundary zoonotic arboviruses that are recognized as emerging animal disease threats. The area of focus will be directed toward diagnostic assays for Japanese encephalitis and Rift Valley fever viruses. Specific objectives include: 1) develop and generate new monoclonal antibodies (mAbs) to Japanese encephalitis virus, 2) develop and validate new competitive enzyme-linked immunosorbent assays (cELISAs) for Japanese encephalitis virus and 3) develop and validate new cELISA for Rift Valley fever virus.
Approach:
The aim of this project is to develop competitive enzyme-linked immunosorbent assays (cELISAs) specific for the transboundary arboviral disease threats Japanese encephalitis virus (JEV) and Rift Valley fever virus (RVFV). Objective 1: The approach will entail first designing recombinant peptides. Recombinant peptides will be used for mouse injections. Test-bleeds from each mouse will be screened to select which animals are producing the most specific antibodies. The selected animals will be used to generate hybridoma clones to produce monoclonal antibodies (mAbs). Each hybridoma clone will be screened to select clones that produce the most specific mAbs. Following clone selection, sub-clones will be generated, which will undergo a final screening for specificity prior to scaling up production of mAbs. Objective 2: The monoclonal antibodies developed in Objective 1 will be used to develop JEV cELISA. The JEV cELISA will be optimized for sensitivity to the multiple genotypes of JEV. In addition, the cELISA will be optimized for specificity to JEV and cross-reactivity to closely related viruses will be examined. Experimental and, if possible, field samples will be used to validate the assay. Objective 3: Monoclonal antibodies were previously developed at BioStone. To determine the best mAB to use in cELISA, these mABs will be validated for specificity to RVFV. The RVFV cELISA will be optimized for sensitivity and specificity to RVFV. In addition, cross-reactivity to closely related viruses will be examined. Experimental and, if possible, field samples will be used to validate the assay.