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    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; 54(Suppl. 1) — S. 644–649
    ISSN: 0005-2086
    Sprache
    Englisch
    Verweise
    Pubmed: 20521708
    Kontakt
    Institut für Geflügelkrankheiten

    Königsweg 63
    14163 Berlin
    Tel.+49 30 838 62676 Fax.+49 30 838 62690
    email:gefluegelkrankheiten@vetmed.fu-berlin

    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.