Journal article
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
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.
Language: | English |
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Publisher: | Frontiers Media S.A. |
Year: | 2019 |
Pages: | 2119 |
ISSN: | 16643224 |
Types: | Journal article |
DOI: | 10.3389/fimmu.2019.02119 |
ORCIDs: | Vendramin, Niccolò |
Antibody Atlantic salmon (Salmo salar L.) Bead-based immunoassay Heart and skeletal muscle inflammation Heat inactivated plasma IgM Piscine orthoreovirus (PRV)
Animals Antibodies, Viral Capsid Proteins Fish Diseases Immunoassay Immunoglobulin M Immunologic diseases. Allergy Orthoreovirus RC581-607 Reoviridae Infections Salmonidae antibody bead-based immunoassay heart and skeletal muscle inflammation heat inactivated plasma sigma 1 protein, reovirus