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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

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Animal Influenza Virus

Part of the book series: Methods in Molecular Biology ((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.

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References

  1. Ciacci-Zanella JR, Vincent AL, Prickett JR et al (2010) Detection of anti-influenza A nucleoprotein antibodies in pigs using a commercial influenza epitope-blocking enzyme-linked immunosorbent assay developed for avian species. J Vet Diagn Investig 22:3–9

    Article  Google Scholar 

  2. Larsen DL, Karasin A, Zuckermann F et al (2000) Systemic and mucosal immune responses to H1N1 influenza virus infection in pigs. Vet Microbiol 74:117–131

    Article  CAS  Google Scholar 

  3. Loving CL, Lager KM, Vincent AL et al (2013) Efficacy in pigs of inactivated and live attenuated influenza virus vaccines against infection and transmission of an emerging H3N2 similar to the 2011–2012 H3N2v. J Virol 87:9895–9903

    Article  CAS  Google Scholar 

  4. Vincent AL, Lager KM, Janke BH et al (2008) Failure of protection and enhanced pneumonia with a US H1N2 swine influenza virus in pigs vaccinated with an inactivated classical swine H1N1 vaccine. Vet Microbiol 126:310–323

    Article  CAS  Google Scholar 

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Correspondence to Phillip C. Gauger .

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

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Gauger, P.C., Vincent, A.L. (2020). Enzyme-Linked Immunosorbent Assay for Detection of Serum or Mucosal Isotype-Specific IgG and IgA Whole-Virus Antibody to Influenza A Virus in Swine. In: Spackman, E. (eds) Animal Influenza Virus. Methods in Molecular Biology, vol 2123. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-0346-8_22

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  • DOI: https://doi.org/10.1007/978-1-0716-0346-8_22

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-0345-1

  • Online ISBN: 978-1-0716-0346-8

  • eBook Packages: Springer Protocols

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