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Methods Mol Biol. 2014;1161:347-53. doi: 10.1007/978-1-4939-0758-8_29.

Antibody secreting cell assay for influenza A virus in swine.

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1
Virus and Prion Research Unit, National Animal Disease Center, U.S. Department of Agriculture, Agricultural Research Service, 1920 Dayton Avenue, Ames, IA, 50010, USA.

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.

PMID:
24899443
DOI:
10.1007/978-1-4939-0758-8_29
[Indexed for MEDLINE]
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