Impact of analytical variability on clinical interpretation of multiplex pneumococcal serology assays

The response to pneumococcal vaccination can be used to assess a patient's humoral immune response to polysaccharide antigens. Multiplex assays measuring serotype-specific levels of pneumococcal antibodies are often used for this purpose, and clinical algorithms have been published to assist in the definition of an adequate immune response. We evaluated whether interlaboratory variability in multiplex pneumococcal serology assays would affect the clinical classification of the immune response. Specimens from 57 patients were analyzed at three reference laboratories with different multiplex assays to measure pneumococcal serology. Analytical correlation and clinical agreement in the classification of a patient's vaccination status by the three methods were compared. Although substantial variation in the quantitative antibody levels measured by different laboratories was seen, the qualitative classification of individual serologic results showed a high degree of agreement between labs and the ultimate classification of a patient as "protected" or "nonprotected" was the same for most patients. The majority of discordant classifications were driven by a systematic bias in results from one of the assays rather than by random error. These data suggest that the use of integrated assessments based on multiple serotypes can compensate for much of the analytical variability seen between laboratories. Knowledge of the analytical performance characteristics of a particular assay is most important when evaluating patients with results near clinical cut points.