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High-Throughput Luminescence-Based Serum Bactericidal Assay Optimization and Characterization to Assess Human Sera Functionality Against Multiple Shigella flexneri Serotypes.
- Source :
-
International journal of molecular sciences [Int J Mol Sci] 2024 Oct 16; Vol. 25 (20). Date of Electronic Publication: 2024 Oct 16. - Publication Year :
- 2024
-
Abstract
- Shigellosis represents a significant global health concern particularly affecting children under 5 years in low- and middle-income countries (LMICs) and is associated with stunting and antimicrobial resistance. There is a critical need for an effective vaccine offering broad protection against the different Shigella serotypes. A correlate of protection has not yet been established but there is a general consensus about the relevant role of anti-O-Antigen-specific IgG and its functionality evaluated by the Serum Bactericidal Assay (SBA). This study aims to characterize a high-throughput luminescence-based SBA (L-SBA) against seven widespread Shigella serotypes. The assay was previously developed and characterized for S. sonnei and S. flexneri 1b, 2a, and 3a and has now been refined and extended to an additional five serotypes ( S. flexneri 4a, 5b, 6, X, and Y). The characterization of the assay with human sera confirmed the repeatability, intermediate precision, and linearity of the assays; both homologous and heterologous specificity were verified as well; finally, limit of detection and quantification were established for all assays. Moreover, different sources of baby rabbit complement showed to have no impact on L-SBA output. The results obtained confirm the possibility of extending the L-SBA to multiple Shigella serotypes, thus enabling analysis of the functional response induced by natural exposure to Shigella in epidemiological studies and the ability of candidate vaccines to elicit cross-functional antibodies able to kill a broad panel of prevalent Shigella serotypes in a complement-mediated fashion.
- Subjects :
- Humans
Dysentery, Bacillary immunology
Dysentery, Bacillary microbiology
Dysentery, Bacillary prevention & control
Dysentery, Bacillary diagnosis
Antibodies, Bacterial immunology
Antibodies, Bacterial blood
High-Throughput Screening Assays methods
Animals
Luminescent Measurements methods
Immunoglobulin G blood
Immunoglobulin G immunology
Luminescence
Rabbits
O Antigens immunology
Blood Bactericidal Activity
Shigella flexneri immunology
Serogroup
Subjects
Details
- Language :
- English
- ISSN :
- 1422-0067
- Volume :
- 25
- Issue :
- 20
- Database :
- MEDLINE
- Journal :
- International journal of molecular sciences
- Publication Type :
- Academic Journal
- Accession number :
- 39456904
- Full Text :
- https://doi.org/10.3390/ijms252011123