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

Escandon, Paulina; Heatley, J Jill; Berghman, Luc R; Tizard, Ian; Musser, Jeffrey MB

The dissimilar results obtained using anti-bird IgY secondary antibody in the Western blot and the dot-blot ELISAs may have been due to multiple factors, including; antigen-antibody reactions, the species used to generate the anti-bird antibody and the different procedural methods of the assays. Many factors influence antigen-antibody reactions, such as pH, temperature, antibody and antigen concentration, and incubation time. 30 In this study, the Western blot procedure allowed for a longer duration of incubation for the antigen-antibody reactions between plasma IgY and the recombinant N-protein and between the anti-IgY secondary antibody and the plasma IgY. The commercially available anti-bird IgY secondary antibody was manufactured using White-crowned sparrow, Ringed turtle dove, domestic chicken, and Muscovy duck IgY and has been reported to react with dove, duck, sparrow, chicken, turkey, emu, ostrich, quail, macaw and cockatiel antiglobulins (Product insert, Bird IgG-heavy and light chain Antibody, Bethyl Laboratories).