ABSTRACT
Seventy-nine suspected false-positive sera, obtained over 1 year from routine submissions for Brucella ovis serological testing, were used in this study. These sera, which exhibited titres in the complement fixation test, but which because of their epidemiological history and their reactions in the enzyme-linked immunosorbent assay and gel diffusion test were suspected to be false positives, were further analysed by immunoblotting. In blots, using B. ovis antigens, rough lipopolysaccharide was identified as the major, immuno-reactive bacterial component. Antibodies against this macromolecule were present in 46.8% of the suspected false-positive sera. In order to find out if rough lipopolysaccharides from other bacterial species could be the possible cause for the suspected false positivity, 23 sera with highest complement fixation titres were reacted in blots with cell extracts from Escherichia coli, Yersinia enterocolitica, Yersinia pseudotuberculosis, Bortedella bronchiseptica, Actinobacillus seminis, Campylobacter fetus fetus, Campylobacter jejuni, Mycobacterium paratuberculosis, Mycobacterium phlei, Corynebacterium pseudotuberculosis and pure lipopolysaccharides from Escherichia coli and Salmonella typhimurium. Despite high frequencies of antibody reaction with proteins in most of these bacterial cell extracts, which reflect the presence of infections with these bacteria, immuno-staining in the rough lipopolysaccharide region was not observed.
ABSTRACT
Six antigen preparations of bovine leukemia virus, including affinity-purified glycoprotein gp51, gradient-purified fetal lamb kidney-bovine leukemia virus antigen, and four crude antigens, were used in combination with several groups of cattle sera, for the evaluation of electrophoretic immunoblotting as a serological test method. Sera (89) from cattle naturally-infected with bovine leukosis virus, a panel of reference sera from infected and uninfected cattle (18), and serial bleedings from experimentally-infected cows (4) were used. Major differences between the six antigen preparations were observed in their reactivity with the various sera. The immunological variabilities of these antigens were confirmed further by their reactions with a gp51-specific monoclonal antibody. The known immunodominant gp51 failed as a reliable indicator for the serological status of the sera in blots when compared to the results on the same sera, two gp51-specific ELISAs and the agar gel immunodiffusion test were used as reference tests. There was a lack of staining of gp51 antigen by many sera, probably due to the labile nature of the gp51 molecule. On the other hand, non-specific staining in the gp51 region appeared with high frequency in some antigens. Antibody staining of the internal viral protein p24 correlated well with the results of the three reference tests. Other bands stained infrequently and were of no diagnostic value.
