Comparison Of Four Anti-Avian IgY Secondary Antibodies Used In Western Blot And Dot-Blot ELISA To Detect Avian Bornavirus Antibodies In Four Different Bird Species

• Published in: Veterinary medicine (Auckland) Vol. 10; pp. 141 - 150
• Main Authors: Escandon, Paulina, Heatley, J Jill, Berghman, Luc R, Tizard, Ian, Musser, Jeffrey Mb
• Format: Journal Article
• Language: English
• Published: New Zealand Taylor & Francis Ltd 01.01.2019; Dove; Dove Medical Press
• Subjects: Antibodies; Antigens; avian ganglioneuritis; E coli; Enzyme-linked immunosorbent assay; Experimental infection; immunodiagnostics; Immunoglobulins; Influenza; Laboratory animals; Nymphicus hollandicus; Original Research; Phosphatase; Polymerase chain reaction; Poultry; Proteins; proventricular dilatation disease; serology; Severe acute respiratory syndrome; Species; Veterinary colleges; Veterinary medicine; West Nile virus; St Louis Missouri; United States > US; Texas; avian ganglioneuritis; immunodiagnostics; proventricular dilatation disease; serology
• ISSN: 2230-2034; 2230-2034
• DOI: 10.2147/vmrr.s227616

This study evaluated the specificity of different avian secondary antibodies used in Western blot and dot-blot ELISA to detect avian bornavirus antibodies in bird plasma. Plasma samples were collected from: two Blue and gold macaws, one positive and one negative for avian bornavirus by RT-PCR; a Cockatiel and a Monk parakeet prior to and following experimental infection; and, two Mallards, one positive and one negative for avian bornavirus by RT-PCR Samples were analyzed by Western blot and dot-blot ELISA that incorporated recombinant avian bornavirus nucleoprotein as the target analyte. Four species-specific anti-IgY secondary antibodies were used in the assays: goat anti-macaw IgY, goat anti-bird IgY, goat anti-duck IgY, and rabbit anti-chicken IgY. In the Western blot, anti-macaw IgY secondary antibody produced strong signals with Blue and gold macaw and Cockatiel positive plasma, but no signal with Mallard positive plasma. Anti-bird IgY secondary antibody produced strong signals with Blue and gold macaw, Cockatiel, and Mallard positive plasma. Anti-duck and anti-chicken IgY secondary antibody produced a strong and moderate signal, respectively, only with Mallard positive plasma. In the dot-blot ELISA, there was a distinct and significant difference ( <0.05) in the signal intensity between the different secondary antibodies within a bird species. Anti-macaw IgY secondary antibody produced significantly ( <0.05) stronger signals than the other secondary antibodies in Blue and gold macaw, Cockatiel, and Monk parakeet positive plasma, while anti-duck IgY secondary antibody produced significantly ( <0.05) stronger signals than the other secondary antibodies in Mallard positive plasma. In testing psittacines with immunoassays, and especially in assays that incorporate short incubation reaction times such as a dot-blot ELISA, species-specific anti-IgY secondary antibodies provided more accurate results.