Detection of Natural Antibodies and Serological Diagnosis of Pneumococcal Pneumonia Using a Bead-Based High-Throughput Assay

Abstract

Surface-exposed proteins of pathogenic bacteria play a critical role during infections . The vast majority of these molecules are able to trigger strong immune responses. Measuring the humoral immune response against pathogenic bacteria through less-time consuming tests is necessary to reduce the window time for the diagnosis of diseases that may be associated with high morbidity and mortality rates. Due to the multiplex setup, Luminex xMAP^® technology allows analysis of immune responses against many antigens in a single assay. Therefore, less volumes of sera samples are needed and inter assay coefficient of variation is much lower in comparison with other immunoassays. With this methodology, the carboxyl groups on the surface of the polystyrene microspheres must first be activated with a carbodiimide derivative prior to coupling antigens . After the antigen is coupled to a microsphere , different microspheres (all having a unique color) can be combined whereafter the presence of specific antibodies directed against the different antigens in sera can be determined simultaneously. The platform here described can also be useful for epidemiological surveillance programs and vaccine studies.