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PubMed1977

Complement "specificity" and interchangeability: measurement of hemolytic complement levels and use of the complement-fixation test with sera from common domesticated animals.

The results from studies to measure lytic complement (C') in sera of different animal species were reviewed. The traditional system, using sheep red blood cells (RBC) and rabbit antibody, was confirmed as the most sensitive to measure C' levels in man, monkey, dog, guinea pig, and rat serums. Sera C' from horse, cow, and sheep were found to be best assayed using rabbit RBC, whereas C' from goat, cat, and rabbit were best assayed with human RBC. Antibodies and C' from the same species usually mediated lysis of foreign RBC, but this lysis occurred more readily with some RBC targets than with others and may be associated with the presence of natural antibodies in the test sera. The effects of the species origin of a C' source in immunologic reactions in vitro and in vivo are discussed.

Animals↗
PubMed1983

Hotting-up the complement-fixation test.

A detailed investigation into the effect of modifying the incubation temperature of the complement-fixation (CF) test is described. For varicella-zoster virus cytomegalovirus and rubella virus, increasing the incubation temperature progressively increased the sensitivity of the CF test to reach a maximum at 15 degrees C, at which temperature the geometric mean titre of seropositive samples was significantly greater than that found at 4 degrees C. For these three viruses, each serum shown to contain IgG antibodies by ultrasensitive radioimmunoassay procedures was detected by CF following incubation at 15 degrees C. No false-positive reactions occurred at 15 degrees C, but it was our impression that anticomplementary activity was enhanced at this temperature. Significant increases in antibody titre at 15 degrees C were also seen when measles virus, respiratory syncytial virus, adenovirus and Mycoplasma pneumoniae were employed as CF antigens. The results demonstrate that the CF test should be performed at 15 degrees C if optimum sensitivity is to be achieved. The ability of the test to detect significant rises in antibody titre was not impaired at the higher incubation temperature.

Adenoviridae↗
PubMed1977

A complement fixation test for the detection of Eperythrozoon infection in sheep.

A method is described for preparation of an antigen from E. ovis-infected blood first lysed by freezing and thawing, and then followed by sonic disruption of the centrifuged deposit. It was then used in a complement fixation test to demonstrate antibodies in sheep infected with E. ovis. The antigen was not anti-complementary and it retained its potency for long periods when stored concentrated, at about -20 degrees C. Repeated freezing and thawing also has had no effect on the potency of the concentrated antigen. The testing of 668 serums from commercial flocks where E. ovis infection has been diagnosed microscopically has shown the CFT to be a valuable diagnostic aid mainly on a flock sample basis, though the test could be of value on an individual sample basis if the sample were taken from a recently infected sheep.

Anaplasmataceae Infections↗
PubMed1996

Attempted definition by immunoblotting of the causes of reactivity in suspected false-positive sera in the Brucella ovis complement fixation test.

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.

Journal Article↗
PubMed1989

Comparison of the dot-immunobinding assay with the complement fixation test for the detection of Brucella antibodies in sheep.

A dot-immunobinding assay (DIA), using as antigen a sonic extract of Brucella abortus dotted on nitrocellulose bound to a plastic strip, was employed for the detection of Brucella antibodies in 666 sheep sera. The results were compared with the complement fixation test (CFT). All the 242 sera belonging to two flocks were found to be negative by DIA. CFT was negative in 239 cases, whereas three samples showed anti-complementary activity. Of the 424 sera from the remaining three flocks, 98 were positive by both tests and six were positive in DIA, but negative in CFT. In addition, 14 of the 19 anti-complementary sera were also positive by DIA.

Animals↗
PubMed2003

Comparison of complement fixation test, immunoblotting, indirect ELISA, and competitive ELISA for detecting antibodies to Mycoplasma mycoides subspecies mycoides small colony (SC) in naturally infected cattle from the 1995 outbreak in Botswana.

Four serological tests were compared in order to evaluate their efficacies in detecting antibodies to M. mycoides subspecies mycoides SC in cattle sera sampled in 1995 from herds affected with contagious bovine pleuropneumonia (CBPP) in the north-western part of Botswana. Tests that were compared included immunoblotting test (IBT), indirect enzyme-linked immunosorbent assay (i-ELISA), competitive enzyme-linked immunosorbent assay (c-ELISA) and complement fixation test (CFT). The percentages of seropositive samples in the iELISA (48%) and in the c-ELISA (48%) were similar but were comparatively lower than those obtained by the IBT (57%) and CFT (61%). The percentages of positive sera in the IBT and CFT were also similar and overall the efficacy of these tests was better than that of the two ELISA tests. There was 95.5% agreement between the IBT and CFT, 85% agreement between the IBT and c-ELISA, 90.9% agreement between the IBT and i-ELISA, 88.6% agreement between the i-ELISA and CFT, 80% agreement between the c-ELISA and CFT and 90% agreement between the two ELISA tests. It became clearly evident from this comparative study that no single serological test was capable of detecting all animals affected by CBPP under natural field conditions of infection.

Animals↗
PubMed1994

Comparison of particle concentration fluorescence immunoassay to card and complement fixation tests using isolation of Brucella abortus as the standard.

Serologic test data and bacteriologic culture results from 816 cattle were evaluated. Brucella abortus (field strain or strain 19) was isolated from 27.3% of the cows. Results of the card test, particle concentration fluorescence immunoassay (PCFIA), and complement fixation (CF) test were compared. Antibody titers were directly associated with B. abortus isolation, with the majority (64.7%) of field strain isolations having a PCFIA value of < or = 0.300 and a CF test result of at least 1+ at a 1:160 dilution. The specificity of the CF test was significantly higher than that of the PCFIA (at CF > or = 1:40 and PCFIA < or = 0.300).

Agglutination Tests↗