title: A Pneumococcal Protein Array as a Platform to Discover Serodiagnostic Antigens Against Infection.
abstract: Pneumonia is one of the most common and severe diseases associated with Streptococcus pneumoniae infections in children and adults. Etiological diagnosis of pneumococcal pneumonia in children is generally challenging because of limitations of diagnostic tests and interference with nasopharyngeal colonizing strains. Serological assays have recently gained interest to overcome some problems found with current diagnostic tests in pediatric pneumococcal pneumonia. To provide insight into this field, we have developed a protein array to screen the antibody response to many antigens simultaneously. Proteins were selected by experimental identification from a collection of 24 highly prevalent pediatric clinical isolates in Spain, using a proteomics approach consisting of "shaving" the cell surface with proteases and further LC/MS/MS analysis. Ninety-five proteins were recombinantly produced and printed on an array. We probed it with a collection of sera from children with pneumococcal pneumonia. From the set of the most seroprevalent antigens, we obtained a clear discriminant response for a group of three proteins (PblB, PulA, and PrtA) in children under 4 years old. We validated the results by ELISA and an immunostrip assay showed the translation to easy-to-use, affordable tests. Thus, the protein array here developed presents a tool for broad use in serodiagnostics.
authors.affiliation: From the ‡Departamento de Bioquímica y Biología Molecular, Universidad de Córdoba; Campus de Excelencia Internacional CeiA3; and Red Española de Investigación en Patología Infecciosa (REIPI), Instituto de Salud Carlos III, Madrid, Spain;
authors.forename: Alfonso
authors.initials: A
authors.lastname: Olaya-Abril
authors.affiliation: From the ‡Departamento de Bioquímica y Biología Molecular, Universidad de Córdoba; Campus de Excelencia Internacional CeiA3; and Red Española de Investigación en Patología Infecciosa (REIPI), Instituto de Salud Carlos III, Madrid, Spain;
authors.forename: Irene
authors.initials: I
authors.lastname: Jiménez-Munguía
authors.affiliation: §Departamento de Sanidad Animal, Universidad de Córdoba; Campus de Excelencia Internacional CeiA3, Córdoba, Spain;
authors.forename: Lidia
authors.initials: L
authors.lastname: Gómez-Gascón
authors.affiliation: ¶Sección de Enfermedades Infecciosas Pediátricas e Inmunopatología, Hospital Universitario Infantil Virgen del Rocío, Sevilla, Spain.
authors.forename: Ignacio
authors.initials: I
authors.lastname: Obando
authors.affiliation: From the ‡Departamento de Bioquímica y Biología Molecular, Universidad de Córdoba; Campus de Excelencia Internacional CeiA3; and Red Española de Investigación en Patología Infecciosa (REIPI), Instituto de Salud Carlos III, Madrid, Spain; mjrodriguez@uco.es.
authors.forename: Manuel J
authors.initials: MJ
authors.lastname: Rodríguez-Ortega
mesh_terms: D000907:Antibodies, Bacterial / Q000097:blood / Q000276:immunology; D000942:Antigens, Bacterial / Q000097:blood / Q000276:immunology; D001426:Bacterial Proteins / Q000097:blood / Q000276:immunology; D002648:Child; D002675:Child, Preschool; D006801:Humans; D007159:Immunologic Tests; D007223:Infant; D011008:Pneumococcal Infections / Q000097:blood / Q000175:diagnosis / Q000276:immunology; D040081:Protein Array Analysis; D040901:Proteomics; D015203:Reproducibility of Results; D012698:Serologic Tests; D013296:Streptococcus pneumoniae / Q000276:immunology*
chemical_list: D000907:Antibodies, Bacterial; D000942:Antigens, Bacterial; D001426:Bacterial Proteins