title: Detection of Staphylococcus aureus cell walls by enzyme-linked immunoassay using antibodies prepared from a semi-synthetic peptidoglycan precursor.
abstract: The peptidoglycan layer of Staphylococcus aureus contains a (Gly)(5) cross-link which is not found in other bacteria, and which could be used to develop a specific immunoassay for detection of S. aureus in MRSA infections. A semi-synthetic route was used to prepare the S. aureus peptidoglycan precursor UDPMurNAc-L-Ala-γ-D-Glu-L-Lys(Gly)(5)-D-Ala-D-Ala, which was covalently attached to carrier protein bovine serum albumin via the UDP nucleotide. Serum raised using this antigen showed specificity for chemically immobilised peptidoglycan monomer containing (Gly)(5), using an ELISA immunoassay. ELISA assays using 0.1 or 1.0 μg samples of cell walls prepared from two MRSA strains and one penicillin-sensitive S. aureus strain, and from three other bacteria, showed the highest response against cell walls containing (Gly)(5), with a particularly high response against cell walls from one MRSA strain. Competition assays to investigate antibody selectivity demonstrated that the antibody response could be most effectively antagonised using ligands containing (Gly)(5). These data demonstrate that it is possible to generate antibodies with high affinity and selectivity for the (Gly)(5) containing monomer in S. aureus peptidoglycan, that could be used to develop an immunoassay for S. aureus.
authors.affiliation: Department of Chemistry, University of Warwick, Coventry, CV4 7AL, UK.
authors.forename: Sandeep
authors.initials: S
authors.lastname: Sandhu
authors.forename: James A
authors.initials: JA
authors.lastname: Schouten
authors.forename: Julie
authors.initials: J
authors.lastname: Thompson
authors.forename: Mark
authors.initials: M
authors.lastname: Davis
authors.forename: Timothy D H
authors.initials: TD
authors.lastname: Bugg
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D002417:Cattle; D002473:Cell Wall / Q000378:metabolism*; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D007106:Immune Sera / Q000276:immunology*; D009842:Oligopeptides / Q000138:chemical synthesis* / Q000737:chemistry / Q000276:immunology*; D010457:Peptidoglycan / Q000737:chemistry*; D013211:Staphylococcus aureus / Q000166:cytology*
chemical_list: D007106:Immune Sera; D009842:Oligopeptides; D010457:Peptidoglycan