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United States Department of Agriculture

Agricultural Research Service

Research Project: INTERVENTION STRATEGIES TO CONTROL VIRAL DISEASES OF SWINE Title: Antibody secreting cell assay for influenza A virus in swine

Authors
item Kitikoon, Pravina
item LOVING, CRYSTAL
item VINCENT, AMY

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. Humana Press. 1161:347-353.

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.

Last Modified: 7/28/2014
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