Morgan, Staci-Ann2025-10-172025-10-172025-10-16https://hdl.handle.net/10222/85501Bovine viral diarrhea negatively impacts animal health and producer profits in beef and dairy herds worldwide. There is no cure for BVDV, so strict disease management and preventative practices are needed to mitigate the adverse effects of the disease. Disease control practices require accessible and affordable BVDV diagnostic antigen AND antibody tests. While BVDV rapid lateral flow antigen tests are widely available, access to POC antibody detection tests remains limited, with poor diagnostic accuracy. Therefore, we developed a vertical flow rapid antibody test, utilizing novel technologies to detect BVDV antibodies in cattle blood. Thus, this project aimed to develop a novel, accurate, and affordable rapid POC test for the detection of BVDV antibodies in bovine blood. Conjugated HRP detectors and recombinant BVDV capture antigens were utilized to develop a vertical flow immunoassay. Optimization testing was performed to determine the best conditions for detecting the target BVDV antibodies, yielding a test that could detect antibody titers as low as 1:4 in bovine plasma in under 10 minutes.Bovine viral diarrhea virus (BVDV) is a highly contagious pestivirus responsible for many production-limiting symptoms in cattle worldwide. There is no cure for BVDV, and management is highly dependent upon preventative care, immunization, and establishment of herd immunity. A point-of-care (POC) BVDV antibody detection test would allow producers to quickly and effectively determine their herd's immunity and infection status, greatly aiding BVDV management programs. Thus, this project aimed to develop a novel, accurate, and affordable rapid POC test for the detection of BVDV antibodies in bovine blood. Conjugated HRP detectors and recombinant BVDV capture antigens were utilized to develop a vertical flow immunoassay. Optimization testing was performed to determine the best conditions for detecting the target BVDV antibodies, yielding a test that could detect antibody titers as low as 1:4 in bovine plasma in under 10 minutes. A total of 195 positive and 52 negative plasma samples from both beef and dairy cattle were run on the vertical flow test (VFT) and a reference industry standard enzyme-linked immunosorbent assay (ELISA) was used to determine the test's diagnostic sensitivity (DSe) and specificity (DSp). The VFT had a DSe of 98.5% and DSp of 96.2%, while the reference ELISA had a DSe of 97.4% and a DSp of 96.2%. Comparatively both tests had substantial agreement (Cohen’s kappa = 0.91). The newly developed VFT has a diagnostic accuracy exceeding other reported rapid antibody diagnostic tests. Furthermore, the VFT’s DSe is comparable to in-lab ELISA results, suggesting that the test could be used to provide highly accurate pen-side results. The shortened performance time, ease of use, and ability of this test to be performed outside of a laboratory could greatly improve BVDV surveillance and management practices worldwide, reducing disease incidence and improving profit outcomes for beef and dairy producers alike.enDiagnosticCattleBVDV