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Comparison of an enzyme-linked immunosorbent assay and a complement-fixation test for the detection of IgG to bovine herpesvirus type 4 (bovine cytomegalovirus).

An indirect ELISA for the detection of bovine immunoglobulin to bovine herpesvirus type 4 (BHV-4) was developed. Three methods of antigen preparation were compared. They included (1) BHV-4-infected Madin Darby bovine kidney (MDBK) cells treated with glycine and then frozen and thawed, (2) BHV-4-infected MDBK cells treated with glycine and then sonicated, and (3) BHV-4-infected MDBK cells treated with a detergent. The antigen preparation that gave the highest reactivity was the first method. We obtained serum samples from 178 cattle in the field and assayed the serum by ELISA and the complement-fixation (CF) test. Eighty-six percent (153) of the serum samples were positive by ELISA, and 70% (124) were positive by the CF test. The ELISA had a higher degree of sensitivity than did the CF test. Also, the ELISA was specific, and the prevalence of BHV-4-infection was more common in beef and dairy herds than previously recognized.

Animals↗

Inactivated-concentrated virus antigen for indirect complement fixation test of foot-and-mouth disease.

To the culture fluids of BHK-21 cells infected with each of types O, A, and Asia 1 of foot-and-mouth disease virus was added acetylethyleneimine to 0.05% (v/v). The mixtures were incubated at 37 degrees C for 24 hours. To them were then added polyethylene glycol 6000 to 10% (w/v), and the mixtures concentrated to one-tenth of the initial volume. The resulting inactivated-concentrated virus antigens showed a complement fixation (CF) titer ranging from 12 to 24. The recovery rate of CF activity was 40 approximately 60%. This activity of each antigen was maintained at 4 degrees C or -70 degrees C for 6 months at least. Experimentally infected cattle were examined for the development of antibody by the aid of the indirect complement fixation (ICF) test with those antigens. As a result, ICF antibody began to be detected 3 approximately 5 days after inoculation. Its titer reached a maximum 10 approximately 14 days after inoculation and decreased gradually thereafter. It was detected even 232 days after inoculation. There was a tendency for the development of ICF antibody to be parallel with that of neutralizing antibody. It was suggested that ICF antibody might be type-specific. In conclusion, the antigens prepared had such high activity that they could be used for the determination of antibody by ICF. In addition, they were of great practical value because of their sufficient keeping quality and safety.

Animals↗

Comparison of an enzyme-linked immunospecific assay (ELISA) with the cold complement fixation test for the serodiagnosis of Brucella ovis infection.

An enzyme-linked immunospecific assay (ELISA) for the serodiagnosis of Brucella ovis infection in sheep is described and compared with the cold complement fixation (CF) test. ELISA was performed in microtiter plates, using horse-radish peroxidase conjugated to anti-normal sheep serum globulins, and hydrogen peroxide plus o-phenylenediamine as substrate. A heated, cell-free B. ovis extract was used as antigen in both tests. ELISA was easier to perform, distinguished better between positive and negative sera, and did not need heat-inactivated sera.

Journal Article↗