A Chromatographic Strip Test for Rapid Detection of One Lineage of the H5 Subtype of Highly Pathogenic Avian Influenza

A rapid immune colloidal gold test strip for detecting 1 lineage of the H5 subtype of highly pathogenic avian influenza was developed based on membrane chromatography with monoclonal antibodies (mAbs) against hemagglutinin (HA) of the H5 subtype of Avian influenza virus (AIV). Bagg albino C mice that were immunized with AIV A/Goose/Guangdong/3/96(H5N1) yielded 4 hybridomas secreting mAbs (1A6, 1B8, 3C4, and 8F6) specific for HA. Monoclonal antibody 3C4 was conjugated with colloidal gold as the detector antibody. On the test strip, 1A6, in combination with rabbit anti-H5N1 polyclonal antibody, was used as the capture complex at the “test line,” and goat antimouse immunoglobulin G antibody was used as the capture antibody at the “control line.” The strip test correctly detected the H5 subtype of AIV in known positive samples and did not react with the standard antigens of AIV H1-H4 or H6-H14, or the antigens of Newcastle disease virus, Infectious bursal disease virus, Infectious bronchitis virus, or Avian i...