Antigen-capture enzyme immunoassay for detection of avian influenza virus in turkeys.
A double-antibody sandwich ELISA (DAS-ELISA) was developed for detection of avian influenza virus (AIV) antigen. A monoclonal antibody to the viral nucleoprotein (NP) was used to coat the ELISA plates. A direct DAS-ELISA and an indirect DAS-ELISA were evaluated. In the direct DAS-ELISA, monoclonal antibody to the AIV NP conjugated with horseradish peroxidase was used. The direct DAS-ELISA was evaluated for its sensitivity to detect purified NP; this procedure detected as little as 0.1 ng. In the indirect DAS-ELISA, rabbit NP antibody and horseradish peroxidase-conjugated goat anti-rabbit immunoglobin were used as primary and secondary antibodies, respectively. The indirect DAS-ELISA was evaluated for its ability to detect the AIV antigen in tracheal and cloacal specimens from turkeys inoculated with AIV. Results of indirect DAS-ELISA were compared with those of conventional virus isolation. Percentage agreement between indirect DAS-ELISA and virus isolation in AIV-positive samples was found to be 76.1% and, in AIV-negative samples, it was found to be 82.1%. These results indicate that the DAS-ELISA might be a viable alternative to virus isolation because of its rapidity, compared with virus isolation.
Kodihalli
S
S
Department of Veterinary PathoBiology, College of Veterinary Medicine, University of Minnesota, St Paul 55108.
Sivanandan
V
V
Nagaraja
K V
KV
Goyal
S M
SM
Halvorson
D A
DA
D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis; D015153:Blotting, Western; D002642:Chick Embryo; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods / Q000662:veterinary*; D009980:Influenza A virus / Q000302:isolation & purification*; D005585:Influenza in Birds / Q000382:microbiology*; D051379:Mice; D008807:Mice, Inbred BALB C; D009698:Nucleoproteins / Q000276:immunology; D011201:Poultry Diseases / Q000382:microbiology*; D012680:Sensitivity and Specificity; D014422:Turkeys / Q000382:microbiology*; D014764:Viral Proteins / Q000276:immunology
D000911:Antibodies, Monoclonal; D009698:Nucleoproteins; D014764:Viral Proteins