[Study of bovine sera by micro-complement fixation test for taxoplasmosis].
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An enzyme-linked immunosorbent assay (ELISA) for specific IgM antibody to Coxiella burnetii was compared with the indirect immunofluorescence (FA) test in studies of 130 serum samples from 38 patients with Q fever. The IgM fractions obtained after rate zonal-density gradient ultracentrifugation of 37 serum samples from 12 patients were also studied in a complement fixation test (CF-DG). Specific IgM antibody to C burnetii was detected by all three methods in sera collected two to eight weeks after the onset of symptoms. The longest period for which specific IgM was shown to persist was 17 weeks by ELISA and FA and 10 weeks by CF-DG. The ELISA is performed with a single dilution of convalescent-phase serum and offers advantages over the subjective FA and the technically tedious CF-DG methods. The estimation of C burnetii-specific IgM by ELISA or FA is useful for the confirmation of infection when a rising titer of complement-fixing antibody cannot be demonstrated because acute-phase blood samples are not available.
The increasing horse trade requires a reliable immunodiagnosis of equine piroplasma infections due to import restrictions imposed by various countries, including the United States of America. It was the aim of our investigations to establish the suitability of serological tests for the detection of parasite carriers and, eventually, to differentiate between Babesia caballi and B. equi infections. The investigations were carried out on 11 ponies with experimentally-induced B. caballi and/or B. equi infection. The infections were confirmed by the demonstration of parasites in blood smears 2-13 days post infection (PI). The complement fixation test (CFT), the indirect immunofluorescence (IIF) and the enzyme-linked immunosorbent assay (ELISA) were employed for the demonstration of antibodies, and different antigen preparations were tested for their suitability. Antibodies could be demonstrated by all three tests. Complement-fixing antibodies disappear after 2-3 months PI in B. caballi-infected horses, while the IIF and ELISA gave positive results during latent infection. A reliable serodiagnosis thus requires the use of the CFT and IIF, since parasite carriers may appear seronegative by the CFT. Serological differentiation between B. caballi and B. equi was possible by CFT and, to a certain extent, by IIF during early infection, but not by ELISA. The successful treatment of B. caballi infections with Berenil could only be confirmed serologically by IIF.
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Enteroviruses were isolated from 27 patients during 1973 and 1974, 21 of these were Coxsackie B, 4 were ECHO and 2 were polio viruses, 5 further viruses could not be identified. Sera from 14 of the Coxsackie patients were investigated twice by means of the Complement Fixation Test (CFT). 11 of them showed a significant rise in antibody titre against one of the 6 different Coxsackie B viruses. No strong correlation was established between the result of the CFT and the isolated type of virus. Nevertheless, the CFT can be helpful in the diagnosis of a Coxsackie B infection. About 200 such cases with varied clinical features were diagnosed in the Department of Virology, Vienna University over the past 3 years by means of the CFT.
Modification of the Microtiter reading mirror used in the standardized diagnostic complement fixation method permits convenient estimation of the results in per cent hemolysis by direct visual comparison with the hemolytic standards.
An enzyme-linked immunosorbent assay (ELISA) was evaluated for detection of antibodies (Ab) against Mycoplasma hyopneumoniae and M. flocculare in sera from swine experimentally infected with these agents. In addition, the ELISA was compared with the complement fixation test (CFT), and radial immunodiffusion enzyme assay (RIDEA) for the demonstration of Ab against M. hyopneumoniae. Twenty two 6-week-old swine from a respiratory disease-free herd were divided into five groups. Two or three pigs from each of the four groups were inoculated, respectively, with M. hyopneumoniae or with M. flocculare while two pigs in each group were contact exposed to the inoculated penmates. A fifth group, consisting of three pigs, served as inoculated controls. Pigs inoculated with M. hyopneumoniae began coughing 13 days post inoculation (PI). Antibodies were first detected 2 weeks PI with the CFT, 3 weeks PI with the ELISA, and 5 weeks PI with the RIDEA. With the ELISA and RIDEA, Ab were still detectable one year PI at a very low level. With the CFT, Ab were not detectable in sera from any swine beyond 5 months PI. At necropsy 1 year PI, no lesions were detected in lungs of any of the animals nor were mycoplasmas detected. M. flocculare inoculated or contact-exposed pigs never evidenced clinical signs. Antibodies against M. flocculare were first detected 5 to 12 weeks PI with CFT, and 6 to 12 weeks PI with the ELISA. Peak optical density (OD) values obtained in the ELISA with M. flocculare Ab were as high as the values obtained with peak M. hyopneumoniae Ab titers. Levels of Ab against M. flocculare were at relatively higher OD at 1 year PI than Ab against M. hyopneumoniae. Sera with high levels of Ab against M. flocculare cross-reacted slightly with M. hyopneumoniae antigen in immunoblotting and ELISA.
In order to evaluate the efficiency of passive hemagglutination (PHA) and complement fixation (CF) tests for the diagnosis of M. pneumoniae infections, 144 serum samples--out of which 81 from patients with respiratory and cutaneous infections and 63 from subjects with other diseases (controls)--were comparatively investigated by the two methods. The PHA test made evident a positivity index of 65% in the case of mixed pneumonias, as against 17% in controls, while the results obtained by CF were almost identical in the two groups investigated. The high specificity and sensitivity of the PHA reaction recommend this method for the diagnosis of acute M. pneumoniae infections.
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Serums from pigs slaughtered at abattoirs were tested for evidence of Mycoplasma hyopneumoniae infection using a complement fixation (CF) test which avoids the procomplementary effect of pig serum. To establish a diagnosis of enzootic pneumonia, the lungs from all sampled pigs were examined for pathological and histological changes consistent with the disease and cultures were made for mycoplasmas and bacteria. The study was carried out at Parkville and Bendigo 160 km apart at different times and all serums were tested at both laboratories. The results agreed closely. Thirty-six of 97 pigs at Parkville and 46 of 99 at Bendigo had enzootic pneumonia. About 80% were positive in the CF test. Sixteen per cent of porkers and 36% of baconers gave false negative reactors, that is, a negative test though lesions were present. About 18% to 36% gave false positive reactions but the level in the porkers in the Bendigo group was significantly higher (p less than 0.02). Possible explanations include, for the false negatives, loss of reactivity caused by circulating antigen and for the false positives, cross reacting antibody produced by another infection or failure to appreciate that lesions of EP were present in lungs because either they were not identified as such or they were not detected. The validity of any serological test for this disease cannot be established while there is a possibility that the present methods used for diagnosis, gross and microscopic examination and recovery of M. hyopneumoniae, fail to detect some infected animals. Other criteria may have to be adopted.
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We present data in support of the use of a heated histoplasmin control for the complement fixation (CF) test for histoplasmosis to assist in the detection of the presence of h or m antibody.
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Evaluation of diagnostic sensitivity and specificity was based on test results of 346 sera from pigs known to be infected and 139 sera from pigs known not to be infected. All sera were tested with a monospecific antigen (serotype 1) and a polyspecific antigen (serotypes 1-5). The sensitivity of the polyspecific antigen was approximately 85% at serum dilution 1:2 and was significantly higher than the monospecific antigen at all serum dilution levels. The specificity of the two antigen preparations was not significantly different at any dilution and increased from approximately 78% to 1:2 to 100% at 1:128. When pigs from herds with unknown incidence of infection were studied, it was found that a high proportion seroconverted, presumably as a response to subclinical infection. However, the antibody titres waned rapidly. This indicated that seroreaction expresses current or recent infection. Thus, the complement fixation test provides a reliable means of diagnosing pleuropneumonia of pigs and might be useful as a tool to control this disease.