We evaluated a multiplexed bead-based assay (xMAP Pneumococcal Immunity assay from Luminex) for the simultaneous determination of antibodies against 14 capsular polysaccharides. Post-vaccination (Pneumovax) antibody concentrations were measured in 35 healthy children, 40 healthy adults, 99 consecutive patients with increased susceptibility to respiratory infection, and 24 patients with a deficient anti-polysaccharide antibody response. The serotype-specific lower 5th percentile (cutoff) value for the post-immunization antibody concentration was determined in healthy individuals. Eleven of 99 patients (11%) failed to mount a response that was >5th percentile of controls for at least 6 of the 14 serotypes tested, whereas only 3 of 75 controls (4%) failed to do so. All patients with known anti-polysaccharide antibody deficiency failed to mount a response that was >5th percentile of controls for at least 6 of the 14 serotypes tested. The XMAP pneumococcal immunity panel appears useful for identifying individuals with a low response to the unconjugated pneumococcal vaccine.