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Enzyme-Linked Immunosorbent Assay for Detection of Serum or Mucosal Isotype-Specific IgG and IgA Whole-Virus Antibody to Influenza A Virus in Swine

  • Phillip C. GaugerEmail author
  • Amy L. Vincent
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 2123)

Abstract

Enzyme-linked immunosorbent assays can be used to detect isotype-specific anti-influenza antibodies in biological samples to characterize the porcine immune response to influenza A virus (IAV). The isotype antibody assay is based on an indirect ELISA using whole influenza virus as antigen and commercial antibodies directed against porcine IgG and IgA. Samples such as serum, nasal wash, and bronchoalveolar lavage fluid allow for evaluation of systemic, upper, and lower respiratory tract mucosal antibody responses, respectively. The isotype ELISA assay is performed in a 96-well format using IAV test antigen and anti-swine IgG or IgA detection antibodies conjugated to an enzyme that catalyze a color change reaction. The optical density of the sample is measured using an automated plate reader. The assay is useful to characterize the IgG or IgA immune response to challenge or vaccination against specific IAV isolates in different compartments of the immune system.

Key words

Influenza A virus Swine ELISA Serum Lavage Nasal wash Isotype Antibody Mucosal Systemic 

References

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Copyright information

© This is a U.S. government work and not under copyright protection in the U.S.; foreign copyright protection may apply. 2020

Authors and Affiliations

  1. 1.Veterinary Diagnostic Laboratory, Department of Veterinary Diagnostic and Production Animal MedicineCollege of Veterinary Medicine, Iowa State UniversityAmesUSA
  2. 2.Virus and Prion Research UnitNational Animal Disease Center, USDA, Agricultural Research ServiceAmesUSA

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