title: Use of synthetic antigens improves detection by enzyme-linked immunosorbent assay of antibodies against abortigenic Chlamydia psittaci in ruminants.
abstract: Synthetic peptide antigens were prepared for use in enzyme-linked immunosorbent assays (ELISAs) to detect serum antibodies against abortigenic strains of Chlamydia psittaci in livestock. Peptide antigens were identified with C. psittaci B577-immune sera by solid-phase scanning of overlapping octapeptides of variable domains (VDs) of the major outer membrane protein of C. psittaci serovar 1 (omp1 type C. psittaci B577). Two VD 4 regions and one VD 2 region were strongly reactive with all C. psittaci B577 antisera. Peptides encompassing these regions were synthesized with biotin and a serine-glycine-serine-glycine spacer at the N terminus and were attached to streptavidin-coated microtiter plates. In direct ELISAs with these plates, the synthetic peptides reacted with C. psittaci B577 antisera, but not with sera from specific-pathogen-free animals. Serum specimens from 40 sheep and 40 cattle, obtained from herds with abortion problems, were screened for antibodies by these C. psittaci B577 peptide ELISAs and an ELISA with recombinant, genus-specific Chlamydia lipopolysaccharide (LPS) antigen. Results from these newly developed ELISAs were compared to those from the reference C. psittaci B577 elementary body (EB) ELISA and the Chlamydia complement fixation test (CFT). The C. psittaci B577 peptide ELISAs, the LPS ELISA, and the EB ELISA correctly identified the presence or absence of antibodies against chlamydiae in all sheep and bovine sera. The Chlamydia CFT, which is the most widely accepted serodiagnostic method for chlamydial infections in animals, correctly identified the presence or absence of antibodies against chlamydiae in only 78 and 4.9% of sheep and bovine sera, respectively. These results suggest that the C. psittaci B577-peptide and Chlamydia LPS ELISAs are superior for the serodiagnosis of ruminant infections with abortigenic chlamydiae, since they are more sensitive than the CFT, they are easy to standardize, and they use readily available synthetic antigens instead of organism-derived CFT antigen.
authors.affiliation: Department of Pathobiology, College of Veterinary Medicine, Auburn University, Alabama 36849-5519, USA. kaltebe@vetmed.auburn.edu
authors.forename: B
authors.initials: B
authors.lastname: Kaltenboeck
authors.forename: D
authors.initials: D
authors.lastname: Heard
authors.forename: F J
authors.initials: FJ
authors.lastname: DeGraves
authors.forename: N
authors.initials: N
authors.lastname: Schmeer
mesh_terms: D000818:Animals; D000907:Antibodies, Bacterial / Q000276:immunology / Q000302:isolation & purification; D000942:Antigens, Bacterial / Q000276:immunology; D001425:Bacterial Outer Membrane Proteins / Q000276:immunology*; D001426:Bacterial Proteins / Q000494:pharmacology; D001710:Biotin / Q000378:metabolism; D002417:Cattle; D002691:Chlamydophila psittaci / Q000276:immunology*; D003168:Complement Fixation Tests; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D000939:Epitopes / Q000276:immunology; D005998:Glycine / Q000378:metabolism; D008070:Lipopolysaccharides / Q000276:immunology; D051379:Mice; D010455:Peptides / Q000138:chemical synthesis* / Q000276:immunology; D009956:Psittacosis / Q000175:diagnosis* / Q000662:veterinary*; D051381:Rats; D011994:Recombinant Proteins / Q000276:immunology; D012680:Sensitivity and Specificity; D012694:Serine / Q000378:metabolism; D012756:Sheep; D013047:Specific Pathogen-Free Organisms; D019809:Streptavidin
chemical_list: D000907:Antibodies, Bacterial; D000942:Antigens, Bacterial; D001425:Bacterial Outer Membrane Proteins; D001426:Bacterial Proteins; D000939:Epitopes; D008070:Lipopolysaccharides; D010455:Peptides; D011994:Recombinant Proteins; D012694:Serine; D001710:Biotin; D019809:Streptavidin; D005998:Glycine