title: A sensitive immunoblotting technique for the serodiagnosis of Brucella ovis infections.
abstract: A simplified electrophoretic immunoblotting technique based on antigen extracted from Brucella ovis cells with sodium dodecyl sulfate/mercaptoethanol was compared with the complement fixation test (CFT), the enzyme-linked immunosorbent assay, and the gel diffusion test. Sera from 89 chronically infected, semen culture-positive rams, 378 sera from B. ovis-infected flocks, 300 sera from accredited disease-free flocks, and 29 sera from specific-pathogen-free sheep were used. The immunoblotting technique had sensitivity and specificity comparable to those of the standard tests and was able to identify several CFT-negative or -borderline sera as positive. The major immunoreactive antigens of B. ovis had molecular masses of 63, 29, 19 kD (proteins) and 8-12 kD (rough lipopolysaccharide). Antibodies against these antigens were present in 96% of CFT-positive sera from infected flocks and in 100% of sera from semen culture-positive rams. However, immunoblotting also identified antibodies to components other than the major antigens in 1% of CFT-negative sera from infected flocks and in 7.7% of the sera from flocks with a history of freedom from the disease. These reactions probably represent cross-reactivities with other microorganisms and were distinguishable from truly positive reactions.
authors.affiliation: Central Animal Health Laboratory, Wallaceville Animal Research Centre, Upper Hutt, New Zealand.
authors.forename: R
authors.initials: R
authors.lastname: Kittelberger
authors.forename: M
authors.initials: M
authors.lastname: Hansen
authors.forename: G P
authors.initials: GP
authors.lastname: Ross
authors.forename: F
authors.initials: F
authors.lastname: Hilbink
mesh_terms: D000818:Animals; D002002:Brucella / Q000302:isolation & purification; D002006:Brucellosis / Q000175:diagnosis / Q000382:microbiology / Q000662:veterinary*; D003168:Complement Fixation Tests / Q000662:veterinary; D004591:Electrophoresis, Polyacrylamide Gel / Q000662:veterinary; D004797:Enzyme-Linked Immunosorbent Assay / Q000662:veterinary; D005260:Female; D015151:Immunoblotting / Q000379:methods / Q000662:veterinary; D008297:Male; D012680:Sensitivity and Specificity; D012756:Sheep; D012757:Sheep Diseases / Q000175:diagnosis* / Q000276:immunology