Detection of Salmonid IgM Specific to the Piscine Orthoreovirus Outer Capsid Spike Protein Sigma 1 Using Lipid-Modified Antigens in a Bead-Based Antibody Detection Assay

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  • Author: Teige, Lena Hammerlund

    Norwegian University of Life Sciences, Norway

  • Author: Kumar, Subramani

    Norwegian University of Life Sciences, Norway

  • Author: Johansen, Grethe M

    Norwegian University of Life Sciences, Norway

  • Author: Wessel, Øystein

    Norwegian University of Life Sciences, Norway

  • Author: Vendramin, Niccolò

    Public Sector Consultancy, National Institute of Aquatic Resources, Technical University of Denmark, Kemitorvet, 2800, Kgs. Lyngby, Denmark

  • Author: Lund, Morten

    Norwegian Veterinary Institute, Norway

  • Author: Rimstad, Espen

    Norwegian University of Life Sciences, Norway

  • Author: Boysen, Preben

    Norwegian University of Life Sciences, Norway

  • Author: Krudtaa Dahle, Maria

    Norwegian Veterinary Institute, Norway

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Bead-based multiplex immunoassays are promising tools for determination of the specific humoral immune response. In this study, we developed a multiplexed bead-based immunoassay for the detection of Atlantic salmon (Salmo salar) antibodies against Piscine orthoreovirus (PRV). Three different genotypes of PRV (PRV-1, PRV-2, and PRV-3) cause disease in farmed salmonids. The PRV outer capsid spike protein σ1 is predicted to be a host receptor binding protein and a target for neutralizing and protective antibodies. While recombinant σ1 performed poorly as an antigen to detect specific antibodies, N-terminal lipid modification of recombinant PRV-1 σ1 enabled sensitive detection of specific IgM in the bead-based assay. The specificity of anti-PRV-1 σ1 antibodies was confirmed by western blotting and pre-adsorption of plasma. Binding of non-specific IgM to beads coated with control antigens also increased after PRV infection, indicating a release of polyreactive antibodies. This non-specific binding was reduced by heat treatment of plasma. The same immunoassay also detected anti-PRV-3 σ1 antibodies from infected rainbow trout. In summary, a refined bead based immunoassay created by N-terminal lipid-modification of the PRV-1 σ1 antigen allowed sensitive detection of anti-PRV-1 and anti-PRV-3 antibodies from salmonids.
Original languageEnglish
Article number2119
JournalFrontiers in Immunology
Volume10
ISSN1664-3224
DOIs
Publication statusPublished - 2019
CitationsWeb of Science® Times Cited: No match on DOI

    Research areas

  • Atlantic salmon (Salmo salar L.), Antibody, IgM, Bead-based immunoassay, Piscine orthoreovirus (PRV), Heart and skeletal muscle inflammation, Heat inactivated plasma

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