A duplex fluorescent microsphere immunoassay for detection of bluetongue and epizootic hemorrhagic disease virus antibodies in cattle sera

Authors: Drolet, Barbara; Reister-Hendricks, Lindsey

Submitted to: Viruses
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 4/13/2021
Publication Date: 4/14/2021
Citation: Drolet, B.S., Reister-Hendricks, L.M. 2021. A duplex fluorescent microsphere immunoassay for detection of bluetongue and epizootic hemorrhagic disease virus antibodies in cattle sera. Viruses. 13(4):682. https://doi.org/10.3390/v13040682.
DOI: https://doi.org/10.3390/v13040682

Interpretive Summary: Bluetongue virus (BTV) and epizootic hemorrhagic disease virus (EHDV) are transboundary, midge-transmitted orbiviruses that cause moderate to severe disease in domestic and wild ruminants. Economic losses to the cattle industry are due to decreased animal production, but most significantly from trade restrictions on movement of animals and animal products. describes a multiplex assay that can detect antibody to either BTV or EHDV or both in a single bovine serum sample. For effective disease surveillance and accurate trade regulation implementation, rapid, sensitive assays are needed that can determine if cattle have been exposed to one or both of these orbiviruses. We describe the development and validation of a bead-based test that simultaneously detects and differentiates antibody to BTV and EHDV from a single bovine serum sample. When compared to tests currently in use for veterinary diagnostic laboratories for orbivirus antibody detection, the bead-based assay showed increased performance.

Technical Abstract: Bluetongue virus (BTV) causes a trans-boundary, internationally reportable disease in cattle, sheep, and white-tailed deer. The closely related, and often co-circulating, epizootic hemorrhagic disease virus causes a clinically similar devastating disease in white-tailed deer, with increasing levels of disease in cattle in the past 10 years. Transmitted by Culicoides biting midges, together, they constitute constant disease threats to the livelihood of livestock owners. Serious economic impacts result from decreased animal production and significant regulatory repercussions. For effective disease surveillance and accurate trade regulation implementation, rapid, sensitive assays that can differentiate exposure of cattle to BTV and/or EHDV are needed. We describe the development and validation of a duplex fluorescent microsphere immunoassay (FMIA) to simultaneously detect and differentiate antibody to BTV and EHDV in a single bovine serum sample. Aggregate performance measurements were higher for the duplex FMIA with simultaneous detection of BTV and EHDV antibodies than commercially available single-plex competitive enzyme-linked immunosorbent assays (cELISA) for each virus antibody separately.