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Comparative studies on the laboratory diagnosis of experimental Trypanosoma congolense infection in sheep.

Comparative studies on the laboratory diagnosis of sheep experimentally infected with T. congolense were carried out by parasitological and serological methods during a period of 53 weeks. Trypanosomes were most numerous in the first weeks after inoculation, when they were detectable by all three parasitological tests: the wet mount preparation, the haematocrit centrifuge technique, and the mouse inoculation test. In the later stages of the infection, trypanosomes were observed intermittently by the HCT until 34 weeks, and were detected by the mouse inoculation throughout the experimental period. The immunolysis test andthe complement fixation test showed positive reactions starting from two weeks up to 30-35 weeks and 40-45 weeks respectively, while the titres of the indirect fluorescent antibody test and the non-specific immunoconglutination tests persisted at a high level throughout the infection. The effect of a trypanocidal drug was best detected by the IK test.

Animals↗

Use of delayed-type hypersensitivity test to diagnose brucellosis in calves born to infected dams.

Calves (n = 2) born to dams with experimentally induced brucellosis, and calves (n = 4) born to dams with naturally occurring infection were examined by the delayed-type hypersensitivity (DTH) test for possible B. abortus infection. The results were compared with the serum agglutination test, complement fixation test, and Coombs test. Five calves were nursed by their dams for 8-10 weeks after birth. One calf was separated from its dam and fed artificial milk. Three to five months after birth, four calves tested seropositive in the serologic tests. Antibodies were detected in one calf as early as 1 week after birth. The calf fed on artificial milk was seronegative 4-5 weeks after birth. All calves reacted to the DTH test antigen from week 12 until the end of the experiment, even though serologic tests were negative. We conclude that the DTH test is a valuable technique for diagnosing Brucella in calves born to infected dams.

Agglutination Tests↗

Prevalence of Brucella pinnipediae in healthy hooded seals (Cystophora cristata) from the North Atlantic Ocean and ringed seals (Phoca hispida) from Svalbard.

Investigations for Brucella-infections were conducted in 29 hooded seals (Cystophora cristata) caught between Svalbard and Greenland (North Atlantic Ocean; Greenland Sea) autumn 2002, and from 20 ringed seals (Phoca hispida) caught in Billefjord, Svalbard, spring 2003. All animals were apparently healthy and were caught in their natural habitat. Bacteriology on tissue samples from ringed seals was negative, whereas Brucella sp. were recovered in tissues from 11 of the 29 hooded seals (38%), with the highest tissue prevalence in spleen (9/29) and lung lymph nodes (9/24). Anti-Brucella antibodies were detected in sera from 9 hooded seals (31%) (EDTA-modified Slow Agglutination test of Wright, Rose Bengal test, Complement Fixation Test, and Protein-A ELISA). The bacterial isolates all belonged to the genus Brucella according to classical biotyping and PCR analysis based on Insertion Sequence IS711, and were shown to be typical marine mammal strains, based on the occurrence of an IS711 element downstream of the bp26 gene. Their dependency on CO2 for growth, and the presence of one copy each of the omp2a and omp2b gene finally classified them as Brucella pinnipediae. Furthermore, all the hooded seal isolates showed an A+ M+ agglutination profile, which is different from the profile of reference seal strain 2/94 (harbour seal, Phoca vitulina). Thus, these results indicate that B. pinnipediae may contain different biovars. The present results suggest that infection with B. pinnipediae is enzootic in this population. Since the hooded seal is commercially hunted and consumed in Norway, the pathological impact of such infections and their zoonotic potential should be further addressed.

Agglutination Tests↗

A review of kala-azar in China from 1949 to 1959.

This is the first account of China as an important area of kala-azar to appear in the west for nearly 50 years. The review is divided into four main sections. The epidemiological aspects which are described include the geographical distribution and endemicity (under the sub-headings natural conditions of the epidemic area, age distribution, sex differences, seasonal changes in incidence) and research on reservoirs (under the sub-headings survey of canine leishmaniasis and survey of other wild and domestic animals). The section on clinical aspects covers symptoms and signs, post kala-azar dermal leishmaniasis, laboratory diagnosis (puncture, serum tests, complement fixation test, intradermal test) and treatment (medication and the effects of treatment). The possibility that two different types of kala-azar occur in China is discussed and the paper concludes with an appraisal of the present position.

Adolescent↗

Antimitochondrial antibodies after acute myocardial infarction.

The development of anti-heart mitochondrial antibodies (AHMA) after acute myocardial infarction (AMI) has been previously demonstrated in experimental studies. We performed a prospective study to check the incidence, variation curve and specificity of AHMA and AMI by using two different immunological tests: complement fixation and antihuman globulin consumption. Serial venous blood samples were drawn from the patients as follows: for the first 14 days after the event, once every other day; from 14 to 60 days after the acute event, once a week. The development of AHMA was demonstrated in the sera of 15/35 (45.4%) patients with AMI by complement fixation test using mitochondrial antigen obtained from normal human heart tissue compared to 12/33 (36.3%) by using mitochondrial antigen from the infarcted region. The antibodies exhibited cross-reaction with mitochondrial antigens obtained from human skeletal muscle, but did not react with heterologous heart mitochondrial antigens. According to the results of the antihuman globulin consumption test, the AHMA is immunoglobulin M (IgM). None of the patients developed postmyocardial infarction syndrome.

Adult↗

Non specific serological reactions in the diagnosis of bovine brucellosis: experimental oral infection of cattle with repeated doses of Yersinia enterocolitica O:9.

Eight heifers were orally infected with 4 x 10(9) colony forming units of a field cattle strain of Yersinia enterocolitica O:9 in a capsule, 5 days a week, for about 9 weeks (day 0-day 64 (D0-D64). The faecal shedding of Y. enterocolitica O:9 began on D5 for seven out of the eight challenged cattle with a high level of excretion during the first month, followed by a decrease till the day of slaughter (D76). Y. enterocolitica O:9 was not isolated from organs collected at slaughter. No clinical symptoms were observed. Hyperplasia of intestinal lymph formations was the sole microscopic lesions observed. Five animals showed a serological reaction against Brucella antigens in at least one of the following tests: Rose-Bengal test, complement fixation test, tube agglutination test or indirect ELISA (iELISA) tests. Only one animal showed a high level of serological response and a positive reaction in the dithiothreitol-microagglutination test. The observed variability in terms of individual sensitivity to the Y. enterocolitica O:9 infection is in agreement with the low individual prevalence rate and the transient serological reaction and faecal Y. entercolitica O:9 shedding observed in herds showing false positive serological reactions in brucellosis.

Agglutination Tests↗

Serologic survey in a colony of captive common marmosets (Callithrix jacchus) after infection with herpes simplex type 1-like virus.

An outbreak of herpesvirus caused the death of four of five common marmosets (Callithrix jacchus) in a private colony. Gross lesions included acute ulcerative gingivitis, glossitis, and enlarged mandibular lymph nodes. Histologically, all fatal cases showed meningoencephalitis and eosinophilia with intranuclear inclusion bodies in neurons and glial cells. A herpes simplex-like virus was cultured from the brain and was identified as herpes simplex type 1 virus or a closely related virus by immunofluorescence. Serologic testing (complement fixation test) indicated that the surviving adult female was serologically positive for more than 4 yr and that the offspring she produced was seronegative. The most likely source of the outbreak was the owner who mouth fed hand-raised offspring.

Animals↗

A serological investigation into contagious caprine pleuropneumonia (CCPP) in Ethiopia.

For a comparison of serological tests for CCPP, sera from 767 goats were examined. They were subjected to three tests: complement fixation test (CFT) with Mycoplasma capricolum subspecies capripneumoniae antigen; blocking ELISA (B-ELISA) with Mycoplasma capricolum subspecies capripneumoniae antigen; and CFT with Mycoplasma mycoides subspecies mycoides small colony type antigen. Antibodies were detected by these three tests in 23%, 2% and 12%, respectively, of sera from districts in which CCPP had not been reported, and in 60%, 83% and 87%, respectively, in sera from areas in which CCPP had been reported. The specificity of the tests is discussed. The use of the B-ELISA test for the diagnosis and for epidemiological studies of CCPP is strongly recommended.

Animals↗

Congenital rubella affecting an infant whose mother had rubella antibodies before conception.

A woman who had had high titres of rubella antibodies some months before she became pregnant gave birth to an infant in whom congenital rubella was confirmed at 4 months. Rubella haemagglutination inhibition tests, complement fixation tests, and immunofluorescence tests with anti-human IgG were carried out on sera from the mother. Rubella antibody titres in sera obtained in March 1971, seven and a half months before conception, were equivalent to 400 units, which is usually taken as indicating good immunity. Rubella virus was isolated from the baby's nose and throat in July 1973 but was not isolated from a cervical swab taken from the mother in December 1973; tests of her immunological competence did not show any definite abnormality. The presence of high levels of rubella haemagglutination inhibition antibodies does not invariably confer immunity or exclude the possibility of congenital rubella in a subsequent pregnancy.

Antibodies, Viral↗

[Dwarfism and the Shereshevskiĭ-Turner syndrome in patients with congenital toxoplasmosis].

Fifty five patients with cerebral and hypophyseal nanism and Shereshevsky-Turner syndrome were examined for toxoplasmosis. The diagnosis of toxoplasmosis was established on the basis of epidemiological and obstetrical anamnesis, clinical and roentgenological data and serological tests (complement fixation test in 2 modifications--by common and droplet method; precipitation test, fluorescent antibody test) and intradermal allergic test with toxoplasmin (ATT). Of 48 patients with cerebral and hypophyseal nanism ATT proved to be positive in 17 (35.4 per cent); it was positive in 3 of 7 patients with Shereshevsky-Turner syndrome. In some of the patients and their mothers serological tests for toxoplasmosis were also positive. Thus, toxoplasmosis infection among the patients examined was 2.5-3 times more incident than the corresponding indices in healthy children. It is supposed that there is pathology of hypothalamic regulation of hypophyseal functions in toxoplasmosis and nanism. A possibility of pathology of the generative apparatus in maternal toxoplasmosis leading to development of chromosomal embryopathies could not be excluded. The authors consider that further studies are necessary for ascertaining (in some of the cases) of the pathogenetic association between toxoplasmosis and growth and developmental disturbances.

Adolescent↗

Antigenic cross-reactions between tick-borne orbiviruses of the Kemerovo serogroup.

Antigenic relationships between the four subgroups of the Kemerovo serogroup of tick-borne orbiviruses were examined. Kemerovo (KEM subgroup), Broadhaven (BRD) and Wexford (WEX) [Great Island (GI) sub-group], Chenuda (CNU sub-group), and Wad Medani (WM sub-group) viruses cross-reacted in immunofluorescence tests. Complement fixation tests (CFT) revealed that KEM virus was more closely related to BRD and WEX viruses than to either CNU or WM viruses. By cross-neutralization, all the viruses were shown to be distinct; only BRD and WEX showed slight cross-reaction. In Vero cells infected with either KEM, BRD, WEX, or CNU viruses, 10 major (p1 to p10) and a variable number of minor virus-induced 35S-labelled polypeptides were detected. Comparison of the polypeptides precipitated by homologous AF revealed close similarities between BRD and WEX viruses but obvious differences between these two viruses and KEM and CNU viruses. In cross-reactions, p6 of KEM and CNU viruses, and p7 of BRD and WEX viruses were immunoprecipitated. Immunofluorescence and immunoprecipitation tests appeared to identify an inter-group specific antigen that was distinct from an intra-group specific antigen detected by CFT. The results support division of KEM-related viruses into 3 new serogroups - KEM (comprising KEM and GI subgroups), CNU and WM - and indicate that a 38 to 43kD polypeptide carries the major inter-group specific antigen.

Animals↗

Immunological studies in human schistosomiasis. III. Immunoglobulin levels, antibodies, and delayed hypersensitivity.

Levels of IgG, IgE, IgM, and IgA were determined, specific antibodies were detected by the fluorescent antibody test, hemagglutination test, complement fixation test and immunoelectrophoresis, and intradermal tests for delayed hypersensitivity to Schistosoma mansoni antigens were performed in Brazilian patients with schistosomiasis mansoni. The results were compared according to the clinical forms of the disease. IgG levels and antibody titers increased progressively in the subclinical, hepatomegalic, and hepatosplenic forms and there was a statistical relationship between IgG levels and the intensity of responses to the four serological tests; Delayed hypersensitivity (DHS) was found more frequently in hepatosplenic patients and more particularly in those with splenomegaly. DHS also correlated with age, but not with sex or with skin color. The strongest DHS reactions were observed in patients 20 to 34 years old, and in those having the highest fecal egg output. IgG levels, antibody titers, and DHS responses decreased after splenectomy and portal filtration of the worms. No significant variation was observed between untreated subjects, patients who were splenectomized and a group not subject to reinfection for 4 yearsk0

Antibodies↗

Evaluation of tests employed in serological diagnosis of brucellosis caused by Brucella ovis.

A survey was carried out to verify the sensitivity and specificity of various tests (complement fixation test--CF; agar gel immunodiffusion--AGID; indirect enzyme linked immunosorbent assay--ELISA; immunoblotting--IB) employed in the serological diagnosis of brucellosis caused by Brucella ovis. The tests were executed on 44 blood serum samples of rams coming from B. ovis-free flocks, 75 of B. ovis experimentally infected rams and 1139 from rams living in flocks where B. ovis had been previously isolated. All tests were performed using B. ovis hot saline extract (HS) as antigen. Sensitivity results were 97.4% for IB, 98.68% for CF, 100% for AGID and ELISA; specificity was 100% for all methods. Concordance values were 89.62% (CF-AGID), 78.77% (CF-ELISA), 77.74% (AGID-ELISA), 65.45% (IB-CF), 62.93% (IB-ELISA), 67.24% (IB-AGID). IB identified antibodies to antigenic components with molecular weight of 67, 63, 58, 55, 38, 35, 32, 30, 28, 25, 23, 21, 20-18 kDa (proteins) and 15-12 kDa (rough lipopolysaccharide).

Animals↗

Antibody response and antigen-specific gamma-interferon profiles of vaccinated and unvaccinated pregnant sheep experimentally infected with Brucella melitensis.

It is well known that the immune response in sheep against Brucella melitensis is subject to individual variation, depending on diverse factors. It bears asking whether these factors (e.g. clinical disease, active infection, state of previous immunity), when affecting a group, can cause variation in the performance of different diagnostic tests. To clarify some of the circumstances in which this immune response can vary, we examine the immune-response profile of sheep protected against the clinical disease by prior vaccination with strain Rev. 1 in comparison with the profile of unprotected females showing the classical brucellosis symptoms. An experimental infection was provoked at midpregnancy under controlled conditions of both non-vaccinated (n=7) and previously Rev.1-vaccinated ewes (n=5). Their immune response was monitored from 7 to 9 weeks before abortion or normal birth to 30 weeks afterwards. Antibody response was assessed by classical tests (Rose Bengal test, complement fixation test (CFT)) in comparison with other diagnostic tests (indirect ELISA (iELISA), competitive ELISA (cELISA), fluorescence polarization assay (FPA), immunocapture test (ICT)). In addition, the cell-mediated immune response was indirectly evaluated by the in vitro antigen-specific release of gamma-interferon. The antibody levels and antigen-specific gamma-IFN profile of the non-vaccinated ewes having the disease and excreting the pathogen was notably high and differed significantly (P<0.05 or P<0.01) from those of vaccinated ewes that neither contracted brucellosis nor excreted the pathogen. In general, all the tests detect the infection in the non-vaccinated ewes with substantial effectiveness. It can be concluded that the high levels of circulating antibodies and of antigen-specific gamma-IFN are related to active Brucella infection. Similarly, the state of protection against the disease, but not necessarily against infection, due to a previous immunization with the Rev. 1 vaccination, appears to be responsible for a low level of detectable immune response. Nevertheless, the design of the study limits conclusions to pregnant ewes and cannot be extrapolated to non-pregnant ewes or rams. Likewise, the study provides no information on animals which are carriers of B. melitensis.

Animals↗

Validation of the fluorescence polarization assay as a serological test for the presumptive diagnosis of porcine brucellosis.

Sera from Canadian pigs (brucellosis free, n = 14037) and sera from pigs infected with Brucella suis (n = 401) were tested by the buffered antigen plate agglutination test, the complement fixation test, an indirect and a competitive enzyme immunoassay and a fluorescence polarization assay. The results were analysed and assay sensitivity and specificity estimates were calculated. The sensitivity and specificity of the tests were as follows: the buffered antigen plate agglutination test, 77.1 and 96.9%; the complement fixation test (considering anticomplementary sera as negative), 93.3 and 95.5%; the complement fixation test (considering anticomplementary sera as positive), 58.1 and 99.9%; the indirect enzyme immunoassay, 94.0 and 97.9%; the competitive enzyme immunoassay, 90.8 and 96.6%; and the fluorescence polarization assay, 93.5 and 97.2%; respectively. It was concluded that the fluorescence polarization assay was a valuable asset to the diagnosis of porcine brucellosis because of its accuracy, ease of performance and relative cost.

Agglutination Tests↗

A comparison of five serological tests for bovine brucellosis.

Five serological assays: the buffered plate antigen test, the standard tube agglutination test, the complement fixation test, the hemolysis-in-gel test and the indirect enzyme immunoassay were diagnostically evaluated. Test data consisted of results from 1208 cattle in brucellosis-free herds, 1578 cattle in reactor herds of unknown infection status and 174 cattle from which Brucella abortus had been cultured. The complement fixation test had the highest specificity in both nonvaccinated and vaccinated cattle. The indirect enzyme immunoassay, if interpreted at a high threshold, also exhibited a high specificity in both groups of cattle. The hemolysis-in-gel test had a very high specificity when used in nonvaccinated cattle but quite a low specificity among vaccinates. With the exception of the complement fixation test, all tests had high sensitivities if interpreted at the minimum threshold. However, the sensitivities of the standard tube agglutination test and indirect enzyme immunoassay, when interpreted at high thresholds were comparable to that of the complement fixation test. A kappa statistic was used to measure the agreement between the various tests. In general the kappa statistics were quite low, suggesting that the various tests may detect different antibody isotypes. There was however, good agreement between the buffered plate antigen test and standard tube agglutination test (the two agglutination tests evaluated) and between the complement fixation test and the indirect enzyme immunoassay when interpreted at a high threshold. With the exception of the buffered plate antigen test, all tests were evaluated as confirmatory tests by estimating their specificity and sensitivity on screening-test positive samples.(ABSTRACT TRUNCATED AT 250 WORDS)

Agglutination Tests↗

[Studies into specificity for dye test (DT), complement fixation (CF), and intradermal test (ST) in diagnosis of toxoplasmosis (author's transl)].

Reference is made to an earlier study into the usefulness of the intradermal test. Now, an account is given of the specificities of tests used at present for the detection of toxoplasmosis, with reference being made to 4,341 intradermal tests, 1,332 dye tests, and 880 complement fixations. -- The three methods are qualitatively and quantitatively compared in great detail, with the close correlations between them explained. Reference is also made to the different origins of the antibody types detectable by the methods concerned. -- It seems to be quite obvious that the production of those antibody types differs in time and site, just as their lives are different in length. These findings actually are used as a point of departure for more detailed methodical proposals that should be followed in the search for an acute infection with Toxoplasma gondii. -- Various effects which might lead to unspecific tests are described in some length. -- All the above studies and findings have been used to compile a diagnosis diagram which may be a good basis for clinical decision-making on therapy.

Complement Fixation Tests↗

[Serological activity of incomplete antibodies].

Blood sera containing incomplete antibodies were studied in various serological tests, by the results of Coombs' and the inhibition of complement fixation tests. It was shown that division of antibodies into complete and incomplete by their serological activity was conditioned. Antibodies detected in Coombs' test could fix the complement. The blocking antibodies depressing the complement fixation test could fix the homologous complement. Proceeding from the latter the author suggests the test of the conglutinating complex fixation which proved to be effective in detection of antibodies inactive in the agglutination and complement fixation tests.

Animals↗