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An immunochemical strategy based on peptidoglycan synthetic peptide epitopes to diagnose Staphylococcus aureus infections
Authors:Carme Pastells  Gerardo Acosta  Nuria Pascual  Fernando Albericio  Miriam Royo  M-Pilar Marco
Institution:1. Nanobiotechnology for Diagnostics (Nb4D), IQAC-CSIC, Jordi Girona, 18-26, 08034 Barcelona, Spain;2. Combinatorial Chemistry Unit, Barcelona Science Park, Baldiri Reixac 10, 08028 Barcelona, Spain;3. Networking Research Center on Bioengineering, Biomaterials and Nanomedicine (CIBER-BBN), Spain;4. Department of Organic Chemistry, University of Barcelona, 080028 Barcelona, Spain;5. School of Chemistry, University of Kwa-Zulu-Natal, 4001 Durban, South Africa
Abstract:The characteristic pentaglycyl cross-bridge of the Staphylococcus aureus peptidoglycan (PG) cell wall component is an attractive epitope to raise specific antibodies against this microorganism. Based on this approach, we report here for the first time a competitive ELISA able to detect S. aureus down to 104 CFU mL−1, without pre-enrichment on cell culture. The antibodies were raised against peptide-protein bioconjugates prepared by covalently coupling peptide haptens (PSau6 and PSau8) designed and synthesized taking into consideration the complex tridimensional structure in the PG polymer. Deglycosylation of the PG under acidic conditions has found to increase assay detectability. Assay performance has been evaluated in clinical samples such as bronchoalveolar lavage (BAL) and bronchoalveolar endotracheal aspirates (BAS) showing promising results for further implementation of this immunoassay as a daily routine diagnostic tool. Cross-reactivity studies have demonstrated that the immunoassay is specific for S. aureus.
Keywords:Staphylococcus aureus  Peptidoglycan  Immunoassay  Diagnostic  Trifluoromethanesulfonic acid  Antibodies
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