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Published on: September 25, 2014
A Pneumococcal Protein Array as a Platform to Discover Serodiagnostic Antigens Against Infection
Alfonso Olaya-Abril1, Irene Jiménez-Munguía1, Lidia Gómez-Gascón2
1From 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;
Insights
Diagnosing childhood pneumococcal pneumonia is difficult. A new protein array identified three key antigens (PblB, PulA, PrtA) for accurate serodiagnosis in children under four.
Area of Science:
- Microbiology
- Immunology
- Proteomics
Background:
- Pneumonia, a severe disease caused by Streptococcus pneumoniae, presents diagnostic challenges in children due to test limitations and distinguishing infection from colonization.
- Current diagnostic methods for pediatric pneumococcal pneumonia are often inadequate, necessitating the development of improved serological assays.
- Serological approaches are gaining traction for diagnosing pediatric pneumococcal pneumonia, offering a promising alternative to existing methods.
Purpose of the Study:
- To develop a novel protein array for simultaneous screening of antibody responses against multiple Streptococcus pneumoniae antigens.
- To identify specific protein antigens that can reliably diagnose pneumococcal pneumonia in children.
- To create a basis for developing user-friendly and affordable diagnostic tests.
Main Methods:
- Proteomics approach involving protease "shaving" of bacterial cell surfaces and LC/MS/MS analysis of 24 pediatric clinical isolates.
- Recombinant production and printing of 95 identified proteins onto an array.
- Probing the protein array with sera from children diagnosed with pneumococcal pneumonia.
Main Results:
- A protein array was successfully developed, enabling simultaneous antibody response screening.
- A discriminant response was identified for three specific antigens (PblB, PulA, and PrtA) in children under 4 years old with pneumococcal pneumonia.
- ELISA and immunostrip assays validated these findings, demonstrating potential for easy-to-use diagnostic tools.
Conclusions:
- The developed protein array serves as a valuable tool for broad serodiagnostic applications in pneumococcal pneumonia.
- The identified antigens (PblB, PulA, PrtA) show promise for accurate and specific diagnosis of pediatric pneumococcal pneumonia.
- This research paves the way for affordable and accessible serodiagnostic tests for children.
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.
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