Identification of Mycoplasma synoviae immunogenic surface proteins and their potential use as antigens in the enzyme-linked immunosorbent assay.
Specific immunogenic proteins of Mycoplasma synoviae (MS) strain WVU-1853 were purified and characterized by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and by Western transfer using anti-MS, anti-M. gallisepticum, and non-immune chicken sera. A cluster of prominent immunoreactive proteins with molecular masses from 46 to 52 kDa (p46-52) and less reactive single proteins of approximately 22 and 92 kDa were shown to partition into the detergent phase of Triton X-114 MS lysates, suggesting that these amphiphilic polypeptides are integral membrane proteins. Monoclonal antibodies, produced by immunizing mice with MS whole cell proteins and shown to bind species-specific determinants, reacted strongly with p46-52 and less intensely with the 22-kDa protein, but they did not react with the 92-kDa protein. A protein fraction extracted from the Triton X-114 detergent phase and further purified by ion-exchange chromatography was found to be highly enriched in p46-52 and was used as antigen in a prototype enzyme-linked immunosorbent assay (ELISA) to detect MS antibodies. Seventy serum samples were taken variously from specific-pathogen-free chickens experimentally infected with MS or MG and from commercial broiler breeder chickens vaccinated for MS and MG. All samples were tested by both rapid serum agglutination and a prototype ELISA described herein; the two tests were strongly correlated (r = 0.776). The results indicate that the ELISA antigen used--the immunodominant cell surface proteins in the p46-52 cluster--appears to be a good candidate for use in the development of improved rapid diagnostics of MS infections in birds.
Gurevich
V A
VA
School of Veterinary Science, University of Melbourne, Parkville, Victoria, Australia.
Ley
D H
DH
Markham
J F
JF
Whithear
K G
KG
Walker
I D
ID
D000818:Animals; D000907:Antibodies, Bacterial / Q000096:biosynthesis; D000911:Antibodies, Monoclonal / Q000096:biosynthesis; D000937:Antigen-Antibody Reactions; D000942:Antigens, Bacterial / Q000276:immunology* / Q000302:isolation & purification; D015153:Blotting, Western; D002645:Chickens; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay; D008565:Membrane Proteins / Q000276:immunology* / Q000302:isolation & purification; D009174:Mycoplasma / Q000276:immunology*
D000907:Antibodies, Bacterial; D000911:Antibodies, Monoclonal; D000942:Antigens, Bacterial; D008565:Membrane Proteins