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Title:   Author  item
                                            GAUGER, PHILLIP - Iowa State Universityitem
                                            Loving, Crystalitem
                                            Baker, Amy
  Submitted to: Methods in Molecular Biology
                                    Publication Type: Book / Chapter
                                    Publication Acceptance Date: 3/10/2014
                                    Publication  Date: 6/30/2014
                                    Citation: Gauger, P.C., Loving, C.L., Vincent, A.L. 2014. 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., editor. Methods in Molecular Biology: Animal Influenza Virus. 2nd edition. Springer New York, Humana Press. 1161:303-312.Interpretive Summary:
                                Technical Abstract: Enzyme-linked immunosorbent assays (ELISA) can be used to detect isotype specific anti-influenza antibodies in biological samples to characterize the porcine immune response to influenza A virus. The isotype antibody assay is based on an indirect ELISA using whole influenza virus as antigen and detection 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 anti-swine 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 response to challenge or vaccination against specific influenza virus isolates in different compartments of the immune system.


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