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Epidemiology and inmunogenetics in recently diagnosed Venezuelan children with insulin-dependent diabetes mellitus.

Genetic and immunological markers in children with Type I diabetes have not been studied previously in Venezuela. We evaluated 91 newly diagnosed IDDM children mean age 7.8 +/- 4.5 (range 0.8-20.8 years), 51 females and 40 males. Eleven percent of first degree relatives had a family history of Type I IDDM; 56.7% had had upper respiratory infection prior to diagnosis and 12.7% had had either mumps or varicella. Peak incidence of disease was found in February and March and August to October. Eighty seven percent had HLA-DR3 and/or DR4 vs 36% of the Venezuelan general population; 81.6% were HLA-DQW2 and/or HLA-DQW8. We found 55.9% to have positive islet cell antibodies (ICA) with 4 of these having a positive complement fixation test. Three patients (7.9%) were found to have positive insulin autoantibodies. Only 3 out of 11 HLA-identical siblings had positive ICAs, while none had positive insulin autoantibodies. One of them also had a positive complement fixation test; this subject developed IDDM. No positive serotypes for enterovirus (Coxsackie-B) were found in our patients, but we detected 11 cases with elevated titers for cytomegalovirus antibodies and positive antibodies for measles, mumps, herpes and varicella were found in some children. These data confirm that most of our Type I diabetics carry HLA-DR3 or DR4 and that the heterozygous DR3/DR4 phenotype is markedly increased in this population; they also indicate that DR3QW2 and DR4QW8 are associated with increased risk in our population.

Adolescent↗

Immunoelectrophoretic analysis of Mycoplasma mycoides var. mycoides.

Acrylamide gel electrophoresis was used to show the similarities and differences in the membrane proteins of two vaccine and two virulent strains of Mycoplasma mycoides var. mycoides. Immunoelectrophoretic (IEP) analysis was also used to partially characterize the associated antigens. Antibody spectra to the antigens of M. mycoides differ in rabbit, pig, and cattle sera. Rabbits produce better precipitating antibody against the anodic migrating protein mycoplasma antigens than cattle and pigs as seen in IEP. However, rabbit anti-M. mycoides serum did not show precipitating antibody against the heat-stable carbohydrate antigen. As judged by IEP, the major carbohydrate antigen extracted from the media, or boiled whole organism, is similar to that present in the sera-infected cattle and knee joints of calves. This carbohydrate antigen has a cathodic migration in IEP at pH 8.6. Periodate oxidation, classically used to destroy carbohydrate, also destroys most of the protein antigens. Heating the antigens to 56 C for 10 min destroys many of the noncarbohydrate antigens and 100 C eliminates all but the carbohydrate antigen. Extraction of M. mycoides with chloroform-methanol, phenol, ethanol, or ethanol-acetone reduced or eliminated most of the protein antigens. Some of the isolated antigenic fractions of M. mycoides were tested to determine their activity in the diagnostic complement fixation test for contagious bovine pleuropneumonia and their inhibitory effect in this test by using bovine anti-M. mycoides antisera having precipitating antibody and circulating antigen. The complement fixation antigen is not the galactan, cannot be extracted by chloroform-methanol, but is stable to boiling at 100 C and may be extracted by phenol and partially precipitated by ethanol-acetone.

Animals↗

Application of the latex agglutination test for human paragonimiasis in South America.

The latex agglutination test (LAT) was compared with the complement fixation test (CFT), double diffusion test (DDT) and enzyme-linked immunosorbent assay (ELISA) for the serodiagnosis of South American paragonimiasis. Sera from 17 Peruvian and Ecuadorian patients with paragonimiasis, from 15 Japanese patients with other helminthiases, and from 100 healthy students were analyzed. The tests were also compared using serum samples from patients treated with bithionol. There was remarkable agreement among the methods and it was concluded that the LAT could be a useful serodiagnostic technique for paragonimiasis, especially in field surveys.

Complement Fixation Tests↗

Epidemiology of bovine anaplasmosis in beef calves in northern Queensland.

Natural infection of calves with Anaplasma marginale was studied in two endemic areas of tropical north Queensland. Infection, assessed by a micro-complement fixation test, occurred throughout the year but was most frequent just after the summer wet season. Infection was clinically inapparent. Calves vaccinated with A. centrale did not show improved growth rate when compared with non-vaccinated calves. The complement fixation test was shown to be most effective in the detection of recent infections.

Anaplasmosis↗

Detection of precipitating antibodies to Histoplasma capsulatum by counterimmunoelectrophoresis.

The technique of counterimmunoelectrophoresis was evaluated for its usefulness in the detection of precipitating antibodies to Histoplasma capsulatum, designated h and m precipitin bands. Forty-four patients with active histoplasmosis had either m bands or both h and m bands. The h precipitin band occurred primarily in patients with disseminated disease, chronic pulmonary disease, or mediastinal lymphadenopathy of several months' duration; with resolution of the infection, this antibody disappeared. The m band appeared earlier in the course of histoplasmosis and persisted for months to years after resolution of the infection. Antibodies detected by counterimmunoeelectrophoresis were in the immunoglobin G class and were more specific for histoplasmosis than those detected by the complement fixation test. Only one false positive h band occurred in a total of 81 sera tested (specimens from 24 healthy control subjects and 57 patients with other diseases). Counterimmunoelectrophoresis was both more sensitive and quicker than immunodiffusion at detecting h and m antibodies. Combining this assay with standard complement fixation tests and fungal culture methods should lead to the accurate, rapid diagnosis of histoplasmosis.

Antibodies, Fungal↗

[The immune response in those ill with and convalescing from Q fever studied by using immunoenzyme analysis].

The study of immune response in Q-fever patients and convalescents at different periods from the beginning of the disease demonstrated the diagnostic possibilities of the complement fixation test (CFT), the microagglutination test (MAT), the passive immunofluorescence test (PIFT) and the enzyme immunoassay (EIA). High effectiveness of EIA in comparison with CFT, MAT and PIFT was noted (97, 63, 87 and 91% respectively). The sensitivity of EIA was not less than 100-250 times higher than that of CFT, 15-100 times higher than that of MAT and 30-80 times higher than that of PIFT.

Agglutination Tests↗

Detection and typing of foot-and-mouth disease virus by enzyme-linked immunosorbent assay: a sensitive, rapid and reliable technique for primary diagnosis.

A highly sensitive indirect sandwich enzyme-linked immunosorbent assay suitable for adoption as the routine diagnostic and typing test for foot-and-mouth disease virus of all seven serotypes is described. The assay uses rabbit and guinea pig antisera raised against inactivated 146S virus antigens. Strong homotypic and minimal heterotypic reactions with both whole virion 146S and derived virion subunit 12S antigens achieved a detection sensitivity approximately 125 times that of the complement fixation test. When applied to diagnostic material with positive-negative threshold criteria generated from testing a large number of negative samples, a positive result was obtained on 83.3 per cent of original virus-positive epithelia, three times the rate for the complement fixation test, and all were typed after one passage in culture.

Animals↗

A passive hemagglutination test for the detection of Brucella infection.

An improved method of coupling a soluble complex, ABS, isolated from B, abortus to sheep red blood cells by chromic chloride in the presence of barbital buffer resulted in a highly specific and reliable passive hemagglutination test for the diagnosis of human and animal brucellosis. No false positive results have been found so far in more than 2000 control sera from known brucellosis-free patients or animals. On the other hand, with over 1800 test sera from patients or animals with Brucella infections, there were no false negative results, in contrast with tube agglutination or complement fixation tests.

Animals↗

Evaluation of the complement fixation and indirect immunofluorescence tests in the early diagnosis of primary Q fever.

A comparison was made of the performance of the commonly used complement fixation test and the more recently developed indirect immunofluorescence test in the early diagnosis of Q fever. The 303 sera tested were from 181 patients who contracted Q fever during an outbreak in Switzerland in 1983. Specific IgM antibodies were detected by the immunofluorescence test in 53% and 89% of sera obtained during the first and second week respectively after onset of illness. With the complement fixation test, the diagnosis could not be made until the second week of illness. The immunofluorescence test proved to be superior to the complement fixation test in the early detection of Q fever. Not only was it more specific but also faster and simpler to perform, permitting an earlier diagnosis on the basis of results obtained with a single serum specimen.

Antigens, Bacterial↗

Plaque assay of Sendai virus in monolayers of a clonal line of porcine kidney cells.

The MN strain of Sendai virus formed distinct plaques in monolayers of PS-Y15 cells, an established porcine kidney cell line. The plaque-forming ability was neutralized by specific antibody to the virus. A linear relationship was found between the concentration of virus and the number of plaques. The sensitivity of this assay was about equal to that of the in ovo titration. When applied to the serum neutralization test, the end points obtained were comparable to those of the hemagglutination-inhibition and complement-fixation tests.

Animals↗

Antigenic similarity of morphologic type CIII1 bacteriophages.

The aim of the study was to demonstrate antigenic differentiation among bacteriophages belonging to the same morphologic type--CIII1 according to Krzywy and Slopek or A2 according to Ackermann. Twenty-six bacteriophages which multiplied on various strains of bacteria of the genera Escherichia, Shigella and Klebsiella, were studied. Serologic tests were done by the quantitative complement fixation test. Immune sera against 5 phi E. coli. D8 Sh. flexneri, G35 Sh. sonnei and Kl7 Klebsiella bacteriophages were obtained from rabbits. It was shown that bacteriophages of CIII1 morphologic type, with morphologically identical virion had different antigenic specificity. On the basis of results obtained the phages could be divided into eight serologic groups. The bacteriophages for which antisera were obtained belonged to four different serologic groups. All bacteriophages had some common antigens with 5 phi E. coli phage. The similarity points to a phylogenetic relationship among phages of CIII1 morphologic type.

Antibodies, Viral↗

[A case of hypersensitivity pneumonitis caused by a humidifier].

A 58-year-old woman was admitted complaining of dry cough and exertional dyspnea. Physical findings, chest X-ray films, chest CT scan and respiratory function tests were suggestive of interstitial pneumonia. Transbronchial lung biopsy showed specific findings of hypersensitivity pneumonitis. As a result of positive provocation test using her home humidifier, a diagnosis of humidifier lung was made. Many microorganisms including Flavobacterium meningosepticum were cultured from the water left in the humidifier for one week. As both complement fixation test and precipitation test were positive to humidifier water and to extract of Flavobacterium meningosepticum, the humidifier and Flavobacterium meningosepticum were suggested to be causative in this case.

Alveolitis, Extrinsic Allergic↗

Platelet aggregation by isolated and aggregated human IgG.

Isolated myeloma proteins of the four human IgG subclasses, aggregated by heat of bis-diazotized benzidine (BDB), induced aggregation of human platelets (Pl.A.) demonstrable by the sedimentation pattern test. With this sensitive technique, approximately 0-1 mug/ml of BDB-aggregated IgG1 and IgG3 could be detected whereas 200 to 1000 times higher concentrations of IgG2 and IgG4 were required for the Pl.A. reaction. The reaction pattern showed similarities with complement fixation tests.

Benzidines↗

Virus serology in familial Crohn disease.

BACKGROUND: Recent findings that early-in-life virus infections represent a risk factor for Crohn disease, that exacerbations of disease sometimes appear associated with common viral infections, and, in particular, suggestions that Crohn disease may be the result of persistent infection with measles virus prompted serologic studies for antibody to 19 common viruses, Chlamydia psittaci, and Mycoplasma pneumoniae. METHODS: Sera from 14 affected members of 2 French families with a high frequency of Crohn disease and from age- and sex-matched controls, taken in 1990 and 1992, and from unaffected family members were tested. Complement fixation, enzyme immunoassay, and indirect immunofluorescent tests were used. RESULTS: There were no significant differences between patients and controls or between affected and unaffected family members of family 2, with the exception that affected siblings carried higher antibody titers for mycoplasma one year and for varicella another year. There were no differences in measles IgM and IgG or mumps IgG antibody levels between patients and controls, or between patients and unaffected family members. CONCLUSIONS: Serology failed to find evidence of participation by 19 common viruses, C. psittaci, and M. pneumoniae in Crohn disease. The data do not support the hypothesis that persistent measles virus infection causes Crohn disease.

Adolescent↗

A solid-phase indirect haemadsorption assay (SPIHA) for detection of immunoglobulin m antibodies to Toxoplasma gondii: application to diagnosis of acute acquired toxoplasmosis.

A solid-phase indirect haemadsorption assay (SPIHA) for the detection of immunoglobulin M (IgM) antibodies to Toxoplasma gondii is described. Polystyrol microtiter plates are coated with anti-human IgM (mu-chain-specific) antibodies and then sequentially allowed to react with patient's serum and sheep erythrocytes sensitized with soluble antigen of Toxoplasma gondii. A total of 111 sera were tested in fluorescent antibody test (FAT and IgM-FAT), complement fixation test (CFT), indirect haemagglutination assay (IHA), and in SPIHA. 47 sera were from individuals with a suspected or verified acute Toxoplasma infection. In most of these cases the SPIHA allowed a clear interpretation with respect to the status of the infection, even when the IgM-FAT was not conclusive. In contrast to IgM-FAT, rheumatoid factor or exceedingly high specific IgG antibodies did not interfere with results in SPIHA. A laboratory-acquired infection enabled us to demonstrate the course of immune response measured by SPIHA as well as by IgM-FAT, FAT, CFT, and IHA. The method proposed here is well appropriated to IgM detection, simple to perform, inexpensive, and thus representing an alternative to IgM-FAT, convenient for the routine laboratory.

Complement Fixation Tests↗

Antimitochondrial antibody profiles in patients with primary biliary cirrhosis before orthotopic liver transplantation and titres of antimitochondrial antibody-subtypes after transplantation.

Four antimitochondrial antibody profiles (A-D) have been defined in primary biliary cirrhosis according to the presence of antibodies to M2, M4, M8, and M9 in ELISA and the complement fixation test: A: anti-M9 positive in ELISA and western blot, B: anti-M9 and/or anti-M2 positive in ELISA, C: anti-M2, -M4 and/or -M8 positive in ELISA, D: anti-M2, -M4, and/or -M8 positive in ELISA and complement fixation test. These profiles predict the outcome of primary biliary cirrhosis in the early stages and reflect differences in the natural course of the disease (benign versus progressive). In this study sera from 29 patients with advanced primary biliary cirrhosis who had received liver transplant were retested before and after orthotopic liver transplantation. Twenty-eight were antimitochondrial antibody/anti-M2 positive, and one patient had only antibodies to nuclear dots in the immunofluorescence test on cell cultures. When the antimitochondrial antibody-profiles in these 28 anti-M2 positive patients were analysed, it became evident that 26 of them belonged to subgroup C or D before orthotopic liver transplantation. Two patients had profile B; one had high titres of antinuclear and smooth muscle antibodies indicating an overlap syndrome between primary biliary cirrhosis and autoimmune chronic active hepatitis. The other patient had antibodies to nuclear dots in association with anti-M2. None of the patients had profile A. Antibody titres were studied after orthotopic liver transplantation in 23 of the 28 patients who survived for 1 to 13 years.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗

[Prognostic significance of antimitochondrial antibody profiles A-D in primary biliary cirrhosis].

Sera from 550 patients with primary biliary cirrhosis were tested by ELISA and the complement-fixation test (CFT) for the antimitochondrial antibodies (AMA) anti-M2, anti-M4, anti-M8 and anti-M9. Four profiles were defined and correlated with morphological and biochemical criteria - A: only anti-M9 positive by ELISA; B: anti-M9 and (or) anti-M2 positive by ELISA; C: anti-M2, anti-M4 and (or) anti-M8 positive by ELISA; D: anti-M2, anti-M4 and (or) anti-M8 positive by ELISA; D: anti-M2, anti-M4 and (or) anti-M8 positive by ELISA and CFT. Liver biopsies were obtained in 385 patients. Analysis of histological stage at the time of first serological testing revealed PBC stages I/II or only nonspecific lesions in all group A patients and in 90% of group B patients, while 49% of patients in group C and 59% of those in group D had stage III/IV or had histological features of chronic active hepatitis or "mixed form". Levels of alkaline phosphatase and GPT were significantly higher in groups C/D than groups A/B patients (P less than 0.05), and bilirubin and IgG levels were more frequently elevated in the former groups of patients (P less than 0.05). It is concluded that classification of PBC into AMA profiles may help in identifying patients at high or low risk of progression of the disease.

Adult↗