<p>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</p>

Abstract

PurposeThis 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.MethodsPlasma 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.ResultsIn 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 (P<0.05) in the signal intensity between the different secondary antibodies within a bird species. Anti-macaw IgY secondary antibody produced significantly (P<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 (P<0.05) stronger signals than the other secondary antibodies in Mallard positive plasma.ConclusionIn 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.

Avian Bornavirus Research—A Comprehensive Review

“…Immunofluorescence antibody assay (IFAT; synonym indirect immunofluorescence assay, iIFA), enzyme-linked immunosorbent assays (ELISA) and WB assays are used for the detection of bornavirus-specific antibodies [ 75 , 91 , 116 , 128 , 212 , 217 , 218 , 224 ]. Cells persistently infected with various bornaviruses can be employed for antibody detection by IFAT [ 75 , 217 ].…”

Purification of Emu IgY for Therapeutic and Diagnostic Use Based on Monoclonal Secondary Antibodies Specific to Emu IgY

“…Secondary Antibody Specific to Emu IgY Secondary antibodies targeting IgY from chicken, 14) duck, 14) turkey, 15) macaw, 14) or pigeon 16) are now commercially available or reported, although the majority of which are the polyclonal products. However, to the best of our knowledge, emu-IgYspecific secondary antibodies are not commercially available.…”