Monoclonal antibody-based sandwich enzyme-linked immunosorbent assay for detection of Bordetella pertussis filamentous hemagglutinin

Abstract

Hybrid cell lines producing monoclonal antibodies against Bordetella pertussis filamentous hemagglutinin (FHA) were established. The specificity of the antibodies was ascertained by enzyme-linked immunosorbent assay (ELISA), sandwich ELISA, and sodium dodecyl sulfate-polyacrylamide gel electrophoresis followed by electroblotting. The monoclonal antibody-based sandwich ELISA was developed for detection of B. pertussis FHA. The assay had a detection limit of B. pertussis FHA in concentrations ranging from 7 to 15 ng/ml. The assay was also able to detect whole B. pertussis, Bordetella parapertussis, and Bordetella bronchiseptica bacteria. No cross-reactions were observed with strains of Branhamella catarrhalis, Neisseria meningitidis, Haemophilus influenzae, Klebsiella pneumoniae, Legionella pneumophila, Streptococcus miteor, or Streptococcus pneumoniae. The monoclonal antibodies might be useful for the detection of soluble antigens and whole bacteria in clinical samples and for studies of the immunochemical structure of B. pertussis FHA.