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Comparison of enzyme-linked immunosorbent assay and complement fixation test for the detection of specific antibody in cattle vaccinated and challenged with Brucella abortus.

An enzyme-linked immunosorbent assay (ELISA) and a complement fixation test (CFT) were applied to sera collected regularly from 60 cattle challenged with Brucella abortus strain 544. Of the 60 cattle, 48 were vaccinated with either B. abortus strain 19 or B. abortus strain 45/20 as calves or adults. The remaining 12 cattle were not vaccinated. Of the 27 sera from cattle found to be infected at slaughter, 9 showed aberrant reactions to the CFT in that a positive titer after challenge was delayed or transient. The performance of the ELISA in these nine cattle with aberrant reactions and the other cattle in the trial was as follows. After vaccination with strain 19 or strain 45/20, the number of weeks at which the ELISA was positive was significantly greater (P less than 0.001) than that for the CFT. The strain 19 vaccine also induced positive responses to the CFT and the ELISA for a significantly longer period (P less than 0.001) than did the strain 45/20 vaccine. For cattle with aberrant reactions, the number of weeks after challenge when the ELISA was positive was significantly greater (P less than 0.001) than that for the CFT. For noninfected cattle, the average number of weeks after challenge when the CFT was negative (35 of 40) was higher than that for the ELISA (18 of 40).

Animals

Alternatives to the standardized laboratory branch complement fixation test for detection of antibodies to Coccidioides immitis.

Antibody titers to Coccidioides immitis, using coccidioidin antigen, were determined by three methods: the standardized Laboratory Branch complement fixation method (LBCF), a modified version of the Viral and Rickettsial Disease Laboratory complement fixation test (VRDL-CF), and a quantitative immunodiffusion test (QID). Of the 133 samples evaluated, 72 were negative by each method and 57 (42 serum samples, 15 cerebrospinal fluid samples) were positive by all three methods. Four additional specimens (1 serum sample, 3 cerebrospinal fluid samples) were positive by QID alone. All positive patients were diagnosed clinically as having pulmonary or extrapulmonary coccidioidomycosis or both. When titers from two methods were compared, the agreement within +/- 1 dilution was VRDL-CF/QID, 88.5%; VRDL-CF/LBCF, 85.2%; and LBCF/QID, 82.0%. The agreement of these methods within +/- 2 dilutions was VRDL-CF/QID, 98.4%; VRDL-CF/LBCF, 96.7%; and LBCF/QID, 93.4%. The VRDL-CF and QID methods are simpler to perform; however, they are yet unrecognized as suitable alternatives to the more cumbersome LBCF. Our data show that they should be considered as options for C. immitis serology.

Antibodies, Fungal

Equine infectious anemia: activity of liquid antigen extracts in the agar-gel immunodiffusion and complement-fixation tests.

Twenty-nine lots of acetone-ether extracted liquid antigen were prepared from the pulp of 11 spleens collected from horses at the acute phase of experimental infection. The lots prepared from the highly reactive pulp resulted in general in a liquid antigen of greater activity than those extracted from weakly reactive pulps. Some variations in activity between lots of antigen prepared from the same spleen were also observed. No matter what the results, given a wide enough variation, all results were reproducible. The procedure permitted production of a greater number of antigen test doses from reactive spleens and rendered usable the spleens which failed to give sufficient reactivity when used as pulp antigen in the agar-gel immunodiffusion test. The activity of each lot of liquid antigen was standardized, first by the complement-fixation test and finally by matching with a reference antiserum in the agar-gel immunodiffusion test.

Animals

Evaluation of five commercial tests: complement fixation, microparticle agglutination, indirect immunofluorescence, enzyme-linked immunosorbent assay and latex agglutination, in comparison to immunoblotting for Mycoplasma pneumoniae serology.

A panel of 68 serum specimens from 41 subjects exhibiting various immunological patterns to Mycoplasma pneumoniae as determined by detection of a 180 kDa protein in immunoblotting was used to compare five commercially available tests based on different methods: complement fixation test (CFT), microparticle agglutination (MAG), indirect immunofluorescence assay (IFA), enzyme-linked immunosorbent assay (Elisa), and latex agglutination (LA). The tests were performed according to the manufacturers' instructions. For the determination of immunity to M pneumoniae, the five tests were in good accordance with immunoblotting: sensitivity was 100% for all the five assays, specificity ranged from 95.6% (MAG) to 82.6% (Elisa) and overall agreement ranged from 98.2% (MAG) to 92.8% (Elisa). The comparisons of antibody rates obtained by the four quantitative tests (CFT, MAG, IFA, Elisa) showed correlation coefficients ranging from 0.87 (CFT-IFA) to 0.67 (CFT-Elisa). Six significant antibody rises demonstrated by immunoblotting patterns were detected by all the tests but Elisa in one case. As a whole, the commercial assays gave satisfactory results for routine determination of immune status to M pneumoniae: CFT was the cheapest test and MAG and LA were the easiest to perform.

Adolescent

[Comparison of the sensitivity of complement-fixation test and latex-fixation test in the demonstration of a migration phase of ascariasis (Ascaris suum) in a non-specific host].

The authors compared the sensitiveness of the complement-fixation reaction and the latex-fixation test in the demonstration of a migration phase of ascariasis in a non-specific host (rabbit). The sensitiveness of the latex-fixation test was higher by the value of one dilution of the geometric series, on an average.

Animals

Comparison of an enzyme-linked immunosorbent assay using monoclonal antibodies and a complement fixation test for cattle vaccinated and infected with Brucella abortus.

A competitive enzyme-linked immunosorbent assay (ELISA) using a monoclonal antibody conjugated to horseradish peroxidase MA(A) and a complement fixation test (CFT) were applied to sera collected over a two-year period from 60 cattle challenged with Brucella abortus strain 544. Forty-eight of the cattle were previously vaccinated with B. abortus strain 19 (S19) or B. abortus strain 45/20 (45/20). After challenge 33 of the cattle remained uninfected and nine of the 27 infected cattle showed aberrant reactions by the CFT. The performance of the MA(A) ELISA was as follows: after vaccination, the MA(A) ELISA, like the CFT, was unable to differentiate infected cattle from those recently vaccinated with S19. After challenge the MA(A) ELISA gave results comparable with the CFT for those cattle with aberrant reactions. For the non-infected cattle there was a similar number of weeks after challenge when both tests were negative. It is suggested that the main advantage of the MA(A) ELISA when compared with the CFT lies in its relatively simple test procedure.

Animals

[Study of the indirect complement fixation test for use in the differentiation of foot-and-mouth disease viruses and the determination of immunity].

Studies were carried out to establish the optimal conditions for the indirect complement-fixation test (ICFT) and explore the possibility to use the test in differentiating the foot-and-mouth disease (FMD) viruses and checking the immunity obtained in survivals or in animals that had been vaccinated against FMD. It was demonstrated that the proper use of ICFT necessitates in turn the use of a minimal amount of antigen, which in the presence of 1-2 E homologous hyperimmune serum completely binds the 1 E complement. Results showed that the sera of survived or vaccinated animals contain specific antibodies of titers of 1:4 up to 1:256. The latter depended on the type of vaccine, the amount of antigen in the vaccinal dose, and the time period from the immunization or survival up to the time of taking the serum samples. It is pointed out that ICFT is a sensitive and specific method for the differentiation of FMD viruses and the study of animals' immunity against FMD.

Animals

Equine infectious anemia: preliminary investigation of the complement-fixation test for the demonstration of antibodies and antigen.

Clinical field cases of equine infectious anemia were studied and the disease was reproduced experimentally in horses. Attempts were made to adapt the complement-fixation test to the detection of antibodies in the serum of infected animals and to the demonstration of antigens in tissue extracts.A moderate complement-fixing antibody response was demonstrated in the serum of horses shortly after primary exposure to the infectious agent. However, this reactivity was of short duration and occurred with normal as well as with infected saline tissue extracts. It was therefore concluded that this reaction was not specific for equine infectious anemia. Possibly it is due to the appearance of auto-tissue antibodies. The value of this reaction in the diagnosis of the infection was limited because of its short duration and absence in chronic infection and following re-exposure to the infectious agent.

Animals

Immunotyping of different strains of Japanese encephalitis virus by antibody-absorption, haemagglutination-inhibition and complement-fixation tests.

The immunological characteristics of 26 strains of Japanese encephalitis virus (JEV) isolated in Japan and Malaya between 1935 and 1966 have been investigated mainly by the antibody-absorption variant of the haemagglutination-inhibition test, and to a certain extent also by conventional haemagglutination-inhibition and complement-fixation tests. The antibody-absorption technique shows promise as a routine method for the immunotyping of JEV.At present, two immunotypes can be distinguished. One comprises 2 strains, Nakayama-NIH and I-58, and is designated as the I-58 immunotype. The other immunotype, JaGAr 01, comprises 17 strains which share the characteristics of the JaGAr 01 strain, including one subline of the Nakayama strain, Nakayama-Yakken. The Nakayama-RFVL strain was found to have the characteristics of both immunotypes. The I-58 immunotype differs more markedly from related arboviruses, such as the Murray Valley encephalitis virus and the West Nile Eg101 strain, than does the JaGAr 01 immunotype.Evidence is presented which suggests that a given JEV strain can change immunotype on repeated passage through mice.

Animals

The effects of IgG2 and of antigen concentration on prozoning in the complement fixation test for bovine brucellosis.

Addition of Brucella-specific IgG2 to solutions of Brucella-specific IgG1 initially induced prozoning and at higher concentrations prevented all reaction in the complement fixation test (CFT) for bovine brucellosis. Some infected cattle may be diagnosed as brucellosis-free due to a high ratio of specific IgG2 to IgG1. Increasing the concentration of antigen in the CFT reduced the tendency to prozone.

Animals

Immune complex decomplementation of canine sera for use in a complement-fixation test for diagnosis of visceral leishmaniasis.

Canine sera frequently become anti-complementary when heat-inactivated at 56 degrees C for 30 min, and generally cannot be used in standard complement-fixation (CF) assays. Therefore, a procedure was developed for decomplementing canine sera by absorption with particulate immune complexes consisting of sheep erythrocyte stroma optimally sensitized with anti-sheep erythrocyte antibody (hemolysin). Canine sera incubated for 20 min at 30 degrees C with sensitized stroma consistently showed less than 10% residual complement and were not anti-complementary. This decomplementation procedure was applied in a complement-fixation (CF) test for detection of serum antibodies during canine visceral leishmaniasis. Two groups of German shepherd dogs were injected intravenously with Leishmania donovani or L. donovani chagasi amastigotes, and the course of infections was followed for 12 weeks. Using freeze-thaw sonicate preparations of L. donovani parasites as antigen, reciprocal CF antibody titers above 24 were detectable in sera 7 weeks after infection and gradually increased to a maximum titer of 775 at 12 weeks. Sera from control dogs had mean titers of 24. This improved methodology enhances the potential of the CF test in the serodiagnosis of canine leishmaniasis.

Animals