Development of a Monoclonal Antibody-based Immunohistochemical Method for Detecting Newcastle Disease Virus

  • YAMAMOTO Yu
    Institute of Animal Health, National Agriculture and Food Research Organization
  • SHIMIZU Shinya
    Institute of Animal Health, National Agriculture and Food Research Organization
  • HIROTA Jiro
    Institute of Animal Health, National Agriculture and Food Research Organization
  • KUROKAWA Aoi
    Institute of Animal Health, National Agriculture and Food Research Organization
  • MASE Masaji
    Institute of Animal Health, National Agriculture and Food Research Organization

抄録

<p>In this study, an immunohistochemical (IHC) technique for detecting Newcastle disease (ND) virus (avian avulavirus 1) using paraffin sections of formalin-fixed tissues was developed by production of novel monoclonal antibodies. Mice were immunized with ND virus isolate APMV1/chicken/Japan/Fukuoka-1/2004 (genotype VII), and 14 mouse hybridoma clones producing IgG antibodies against the ND virus were established. Among these antibodies, only three reacted with the viral antigen on the paraffin sections, while one (22C483) exhibited the highest sensitivity in IHC analysis. Results of immunoprecipitation, western blot analysis, and indirect enzyme-linked immunosorbent assay showed that 22C483 reacted with the nucleoprotein of ND virus. Thus, IHC analysis using 22C483, in combination with enzyme digestion treatment for antigen retrieval, can be used for elucidating the pathogenesis of ND.</p>

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