Development of a monoclonal antibody-based competitive inhibition enzyme-linked immunosorbent assay for detection of Bacillus piliformis isolate-specific antibodies in laboratory animals.

A competitive inhibition enzyme-linked immunosorbent assay (ELISA) was developed to detect Bacillus piliformis isolate-specific antibodies in serum specimens from rats and gerbils experimentally infected with B. piliformis isolates R1, R2, or M. Detection was based on the ability of serum antibodies to block binding of B. piliformis isolate-specific monoclonal antibodies to purified B. piliformis flagella. Application of this assay to serum specimens collected from sham-infected or experimentally infected rats and gerbils demonstrated that the serum specimens were capable of specifically inhibiting the binding of B. piliformis isolate-specific monoclonal antibodies to homologous flagella preparations (> 70% inhibition) only when the serum specimens were from animals infected with the homologous B. piliformis isolate. Only one false-negative and false-positive result were obtained when 80 serum specimens were tested by this competitive inhibition ELISA. In addition, we demonstrated that little nonspecific inhibition of monoclonal antibody binding occurred (< 30% inhibition) in this immunoassay specific inhibition of monoclonal antibody binding by serum was due to serum antibody and a serum's ability to inhibit binding of monoclonal antibodies to purified B. piliformis flagella was correlated with antibody reactivity with B. piliformis flagella but not with serum antibody reactivity to whole B. piliformis organisms. These results suggest that this monoclonal antibody-based competitive inhibition assay could be successfully applied to the serologic identification of isolates involved in naturally occurring B. piliformis infections in laboratory animals.