Submitted to: Methods in Molecular Biology
Publication Type: Book / Chapter
Publication Acceptance Date: March 10, 2014
Publication Date: June 30, 2014
Citation: Kitikoon, P., Loving, C.L., Vincent, A.L. 2014. Antibody secreting cell assay for influenza A virus in swine. In: Spackman, E., editor. Methods in Molecular Biology, Animal Influenza Virus. 2nd edition. New York, NY: Springer. p. 347-353. Technical Abstract: An ELISPOT assay to enumerate B-cells producing antibodies specific to a given antigen, also known as an antibody secreting cell (ASC) assay, was adapted to detect B-cells specific for influenza A virus (IAV). The assay is performed ex vivo and enumerates ASC at a single cell level. A simple ASC detection method is based on a solid phase immunoenzymatic principle. The ELISPOT plate is coated with IAV prior to incubation with cell samples. The spot-forming cells are detected by enzyme labeled antibodies directed to the swine immunoglobulin (Ig) class of interest and visualized with the addition of substrate.