A simple and sensitive enzyme immunoassay for determination of soluble type-specific polysaccharide from group B...

A method was developed for the determination of soluble group B streptococcal type-specific polysaccharide in diluted culture supernatant. The type-specific antigen was immobilized to the solid phase of an enzyme immunoassay using wheat germ agglutinin as a link between the plastic surface and the polysaccharide. The binding of monovalent rabbit antiserum to the type-specific polysaccharide was quantitated by an anti-rabbit IgG-enzyme conjugate. Antisera against each of the polysaccharide types Ia, Ib. II and III gave strong and specific reactions against the respective antigen. The ultimate sensitivity of the assay was 70 pg/ml, as determined for type III polysaccharide. Using this technique, it was found that the concentration of soluble type-specific antigen in a GBS, type III culture supernatant reached a steady-state approximately 3 h after the beginning of the stationary growth phase of the bacteria. Ten type III strains were investigated for synthesis of type-specific polysaccharide, and a positive correlation between the production of capsular and soluble type III antigen was found. There was also an inverse correlation between soluble antigen production and the buoyant densities of these strains. The method described may be used for the serotyping of encapsulated GBS and/or determination of soluble type-specific antigen synthesis.