An indirect immunofluorescence (IF) assay has been developed as a useful semiquantitative method for determination of type-specific IgG antibody in human sera to the five serotypes of group B Streptococcus. Antibody titers measured by IF correlated with passive protection in chick embryos, and antibody titers associated with chick embryo protection were delineated. Except for types Ia and Ic, IF antibody to each of the streptococcal types was completely absorbed by homologous strains, and antibody titers were unchanged by incubation with heterologous bacteria. For types Ia and Ic, IF antibody was absorbed by either the Ia or the Ic strain and by native Ia carbohydrate antigen. Antibody titers measured by IF and chick embryo protection against types Ia and Ic were similar, but were divergent for Ib and Ic, a finding suggesting that antibody is predominantly directed to the major carbohydrate determinants. In addition, 29 of 31 sera that had been tested in chick embryos yielded comparable results in mice against challenge with type Ia group B Streptococcus, a finding further validating the chick embryo assay. Sera from all of 43 mothers of infants infected with group B streptococci had antibody titers by IF that were less than titers associated with protection in chick embryos.