Detection of serotype-specific antibodies or capsular antigen of Actinobacillus pleuropneumoniae by a double-label radioimmunoassay

Abstract

Diagnostic tests for Actinobacillus pleuropneumoniae have been problematic because current tests do not use a purified antigen and in most cases measure either antibody or antigen, but not both. We describe a Farr-type double-label radioimmunoassay that utilizes purified, serotype-specific, 3H-capsule to measure antibody to capsule directly or that can measure capsule in a sample indirectly by inhibition of antibody binding. The assay could detect about 1 ng of serotype-specific antibody in serum or at least 100 pg of capsule in a sample. Due to the sensitivity of the assay, false-positive results were common with neat sera (probably due to cross-reacting antibodies to unrelated antigens), but the specificity was improved when the sera were diluted 1:100. The radioimmunoassay should prove to be a useful reference method for research and diagnostic testing and for comparison of new assays for detection of capsule or antibodies to capsule.