ARS | Publication request: Evaluation of a commercial enzyme-linked immunosorbent assay for detection of antibodies against the H5 subtype of Influenza A virus

Submitted to: Journal of Veterinary Diagnostic Investigation
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: November 26, 2012
Publication Date: N/A

Interpretive Summary: Testing for antibodies to Influenza A (IA) virus is a common diagnostic tool used in poultry and also has recently been incorporated into wild bird surveillance efforts. These assays usually are based on the detection of IA nucleoprotein antibodies. Several commercial enzyme-linked immunosorbent assays (ELISA’s) have recently been developed for rapid testing of subtype-specific influenza A (IA) virus antibody in wild birds and poultry, but studies validating their use are limited. In this study, the ID Screen Influenza H5 Antibody Competition ELISA was tested for the detection of antibodies to the H5 subtype of IA virus in waterfowl. Sera obtained from Mallards (Anas platyrhynchos) and Pekin Ducks (Anas platyrhynchos domestica), experimentally infected with low pathogenic (LP) or highly pathogenic (HP) H5 IA viruses, were tested. Three incubation periods (1, 4 and 18 hr) and two dilutions (1:2 and 1:5) were used. Overall, the ID Screen Influenza H5 Antibody Competition ELISA provides a potentially valuable screening tool for the detection of antibodies directed against the H5 subtype of IA virus in waterfowls.

Technical Abstract: Serologic tools for rapid testing of subtype-specific influenza A (IA) virus antibody in wild birds and poultry are limited. In the current study, the ID Screen Influenza H5 Antibody Competition enzyme-linked immunosorbent assay (ELISA) was tested for the detection of antibodies to the H5 subtype of IA virus in waterfowl. Sera obtained from Mallards (Anas platyrhynchos) and Pekin Ducks (Anas platyrhynchos domestica), experimentally infected with eight low pathogenic (LP) and nine highly pathogenic (HP) H5 IA viral strains, were tested. Assays were conducted using three incubation periods (1, 4 and 18 hr) and two dilutions (1:2 and 1:5). The 18 hr incubation and 1:2 dilution provided the best results. All serum samples from LP H5 infected birds tested positive; however, improved detection rates were observed for viruses belonging to the H5N1 clade 2.2.1 as compared to those belonging to clade 2.1.3. Overall, based on results from the modified protocol, the ID Screen Influenza H5 Antibody Competition ELISA provides a potentially valuable screening tool for the detection of antibodies directed against the H5 subtype of IA virus in waterfowls.