Publikationsdatenbank

Production of H5-specific monoclonal antibodies and the development of a competitive enzyme-linked immunosorbent assay for detection of H5 antibodies in multiple species (2010)

Art

Zeitschriftenartikel / wissenschaftlicher Beitrag

Autoren

Dlugolenski, D.

Hauck, R.

Hogan, R. J.

Michel, F.

Mundt, E.

Quelle

Avian diseases

Bandzählung: 54

Heftzählung: Suppl. 1

Seiten: 644 – 649

ISSN: 0005-2086

Sprache

Englisch

Verweise

Pubmed: 20521708

Kontakt

Nutztierklinik: Abteilung Geflügel

Königsweg 63
14163 Berlin
+49 30 838 62676
gefluegelkrankheiten@vetmed.fu-berlin.de

Abstract / Zusammenfassung

The hemagglutinin gene of an avian influenza virus (AIV) A/duck/NC/674964/07 (H5N2) was cloned and expressed in a baculovirus system (H5-Bac). In parallel, a recombinant hemagglutinin of A/Vietnam/1203/04 (H5N1) was expressed in mammalian cells, purified, and used for generation of H5-specific monoclonal antibodies (MAb). The purified H5-Bac was used to develop a competitive enzyme-linked immunosorbent assay (cELISA) to detect H5 antibodies in a species-independent approach using one of the established H5-specific MAbs as the competitor antibody. The cELISA performed with influenza antibody-free sera or with sera of animals infected with other than H5-encoding AIV showed no significant inhibition of H5-MAb binding, indicating high test specificity. In contrast, sera of poultry (chickens, turkeys, ducks) experimentally infected with H5-encoding AIV were able to significantly inhibit the binding of the MAb in a species-independent approach. Comparison of the results of the cELISA with results obtained by a hemagglutination inhibition assay showed a gradient of the sensitivity (turkeys > ducks > chicken). The described results show that H5-specific antibodies in sera can be detected in a species-independent approach by using a recombinant protein.