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ARS Home » Midwest Area » Ames, Iowa » National Animal Disease Center » Virus and Prion Research » Research » Publications at this Location » Publication #300103
Title: Antibody secreting cell assay for influenza A virus in swine

Author
item Kitikoon, Pravina
item Loving, Crystal
item Baker, Amy

Submitted to: Methods in Molecular Biology
Publication Type: Book / Chapter
Publication Acceptance Date: 3/10/2014
Publication Date: 6/30/2014
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.

Interpretive Summary:

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.