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PubMed1985

Survey of brucellosis in goats and sheep in the Yemen Arab Republic: comparison of tests for Brucella melitensis infection in sheep.

Sera from 538 Yemeni goats and 690 Yemeni sheep were screened for brucellosis by the Rose Bengal Plate Test (RBPT) and reactors confirmed by the complement fixation test (CFT) and the serum agglutination test (SAT). The prevalence among goats was 0.4% and among sheep 0.6%. The prevalence among 183 imported goats and sheep was 4.4%. The sensitivity and specificity of three serological tests available for the diagnosis of brucellosis -CFT, RBPT and SAT - were compared using ovine sera obtained throughout an outbreak of abortion due to Brucella melitensis. The RBPT and the SAT were relatively insensitive compared with the CFT (71 and 44% respectively) and the RBPT was as specific as the SAT when suspicious sera were included. The results suggests that the SAT adds little information when used with other tests and the RBPT has limited applications as a screening test for ovine brucellosis.

Agglutination Tests↗
PubMed1998

[Electrophoretic and immunologic comparative analysis of Mycoplasma pneumoniae and Mycoplasma genitalium proteins].

The Mycoplasma pneumoniae FH strain routinely used in our laboratory for over 25 years as antigen in serological tests, 2 reference M. pneumoniae strains from ATCC (29342 and M129) and 3 isolates of M. pneumoniae obtained in 1995 from pneumonia patients were compared by SDS-PAGE, complement fixation test (CFT) and by Western-immunoblotting against human and rabbit serum samples with high level of mycoplasmal antibodies. On SDS-PAGE all M. pneumoniae strains showed the same number of 23 polypeptides on the gel with identical molecular weights. The same strains on immunoblotting against human and rabbit serum samples showed six bands: 170, 89, 75, 55, 38 and 33 kDa with the strongest antibody staining in 170-(P1 protein) and 89-kDa bands. Because of its known antigenic relationships Mycoplasma genitalium was used for comparison. The pattern of M. genitalium proteins on SDS-PAGE was similar to pattern of M. pneumoniae but distinguishable. On immunoblotting six proteins of M. genitalium (135, 127, 110, 95, 75 and 45 kDa) reacted with human and rabbits immunoglobulins for M. pneumoniae antigens. Furthermore in complement fixation test both antigens, prepared from M. pneumoniae and M. genitalium, reacted as well with human and rabbit immunoglobulins for M. pneumoniae and with rabbit immunoglobulins for M. genitalium. These cross-reactions observed in serological techniques could give false positive results in routine diagnosis of M. pneumoniae infections. In such situations showing on immunoblott of presence in tested serum sample of antibodies to 170- and 89 kDa proteins could confirm M. pneumoniae infection.

Animals↗
PubMed1981

Precipitating carbohydrate antigens of Bacteroides fragilis NCTC 9343.

An antigen fraction isolated by various forms of chromatography from phenol/water extracts of Bacteroides fragilis NCTC 9343 was devoid of phosphorus, fatty acids and amino acids. Rhamnose, fucose, galactose, glucose and a neutral sugar tentatively identified as arabinose were present at molar ratios of 1:1:1.2:9.0.9:0.7. The fraction contained an unidentified reducing and ninhydrin positive substance and trace amounts of glucosamine and galactosamine. The antigen fraction produced three immunoelectrophoretic bands against NCTC 9343 antiserum, and reacted in complement fixation tests.

Antigens, Bacterial↗
PubMed1988

Complement-fixing gastric parietal cell autoantibodies. A good marker for the identification of type A chronic atrophic gastritis.

Using an indirect immunofluorescence (IIF) technique, gastric parietal cell autoantibodies of IgG class (GPCA-IgG) were found in 2% of a normal population, in 5-26% of organ-specific autoimmune subjects and in 100% of patients with pernicious anaemia. With the exception of subjects with alopecia, there was a significantly increased prevalence of GPCA-IgG in autoimmune patients with respect to normal controls. GPCA of IgA class were detected in 22% of GPCA-IgG positive subjects, whereas GPCA of IgM class were uncommon. One-hundred and fifteen subjects underwent gastroscopy and body mucosal biopsy. Histopathological findings of chronic atrophic gastritis (CAG) were present in 71% of GPCA-IgG positive autoimmune patients without pernicious anaemia, in 100% of GPCA-IgG positive patients with pernicious anaemia, and in 20% of GPCA negative autoimmune patients. Complement-fixation test was performed in 46 GPCA-IgG positive subjects without pernicious anaemia using the IIF method. Twenty-nine patients (63%) were found to fix complement fractions till C9 (CF-GPCA) together with properdin factor, and in 25 of them (86%) the histological examination of body gastric mucosa disclosed a CAG (P = 0.0003 versus GPCA-IgG positive/CF negative controls). No significant difference was observed for the prevalence of CAG in GPCA-IgG positive/CF negative subjects with respect to GPCA-IgG negative control group. We conclude that the presence of CF-GPCA represents a useful immunological marker in the identification of CAG, while no predictive value seems to be associated with non-complement fixing GPCA-IgG.

Adolescent↗
PubMed1980

Bovine paratuberculosis I. A herd study using complement fixation and intradermal tests.

A dairy herd (102 cattle) which had been enrolled under a paratuberculosis control program for two years utilizing a complement fixation test (carbohydrate antigen) and intradermal skin test (johnin PPD) was subjected to two further herd tests and followed to slaughter to determine infection status by culture and histology. Mycobacterium paratuberculosis infection was demonstrated in 37 of the animals of which only five were considered reactors on the basis of the last two herd tests applied. Cultural and histopathological evaluation indicated the testing procedures had eliminated heavily infected animals. The limitations of these testing procedures under free stall housing conditions are discussed.

Animals↗
PubMed1976

The in vitro assessment of anti-human lymphocyte globulin.

A simple reproducible in vitro test of anti-human lymphocyte globulin activity is discribed. The method is a "microtitre" complement-fixation test using human platelets as a source of antigen, guinea-pig complement and an amboceptor of sheep red blood cells sensitized by rabbit anti-sheep haemolysin.

Animals↗
PubMedDate not supplied

Subtypes of antimitochondrial antibodies in primary biliary cirrhosis before and after orthotopic liver transplantation.

Antimitochondrial antibodies are markers for primary biliary cirrhosis and probably reflect a specific defect in immunoregulation underlying this disease. Antimitochondrial antibodies and their primary biliary cirrhosis-specific subtypes were tested before and up to 6 years after orthotopic liver transplantation. Sera from 31 consecutive patients were tested, 15 patients had primary biliary cirrhosis and 16 non-primary biliary cirrhosis. Antimitochondrial antibodies were investigated under code by immunofluorescence, and primary biliary cirrhosis-specific subtypes were determined by radioimmunoassay (anti-p62, anti-p48) and complement fixation test (anti-M2, anti-M4, anti-M8). Before orthotopic liver transplantation, antimitochondrial antibodies were detected by immunofluorescence in 13 of 15 patients with primary biliary cirrhosis. Of these patients, 12 were positive for anti-p62 and 8 for anti-p48. Ten patients were positive for anti-M2, 4 patients for anti-M4 and 7 patients for anti-M8. Two primary biliary cirrhosis patients and all non-primary biliary cirrhosis patients were negative with all tests. One month after orthotopic liver transplantation, antimitochondrial antibodies titers declined or became negative by antimitochondrial antibodies immunofluorescence, 3 patients became negative by radioimmunoassay for anti-p62 and 1 for anti-p48. With complement fixation test, 4 patients became negative for anti-M2, 2 for anti-M4 and 4 for anti-M8. Antimitochondrial antibody titer reduction observed 1 month after orthotopic liver transplantation remained unchanged in most sera during the following years. A rise was observed in two patients after 4 and 5 years.(ABSTRACT TRUNCATED AT 250 WORDS)

Adolescent↗
PubMed1984

Experimental infection of castrated lambs with Mycoplasma agalactiae.

The course of experimental infection in groups of 6-month-old castrated lambs with field isolates of Mycoplasma agalactiae from France was followed culturally and serologically for 7 months. Infection with an ovine field isolate following inoculation by different routes and contact exposure was compared with that caused similarly by a caprine field isolate. The prolonged infections produced were symptomless apart from limited arthritis in one animal inoculated with the isolate from sheep and increased lachrymation in another associated with the goat isolate. The ovine isolate was more virulent in that ante- and post-mortem recoveries of the organism were more consistent and the serological responses more pronounced. Serological responses varied between animals and between strain infections, and the results of the film inhibition test were more consistent than those of the complement fixation test. The limitations of both these tests for detecting carrier infections are discussed.

Animals↗
PubMed1975

Characterization of autoantigenic sites on isolated dog heart mitochondria.

1. Anti-heart mitochondria autoantibodies were developed in serum from dogs following experimental myocardial infarction. 2. Heart mitochondria frozen and thawed repeatedly in a sucrose/Tris-chloride buffer retained both their functional integrity as measured by the respiratory control ratio and their ability to serve as an antigen in a complement fixation test. Mitochondria frozen and thawed in a potassium chloride/Tris-chloride buffer lost both their functional integrity and their autoantigenic activity after one freeze-thaw cycle. 3. Extraction of the heart mitochondria with acetone/water mixtures to remove phospholipids from the membrane led to a complete loss of the ability of the mitochondria to react in the complement fixation test but did not affect the ability of the membranes to bind autoantibody in absorption experiments. 4. Treatment of the mitochondrial membranes with increasing concentrations of trypsin caused a loss of up to approximately 50% of the membrane protein with a gradual decrease in the autoantigenic activity of the membrane without impairment of the ability of the membrane to bind autoantibody. 5. Removal of up to 90% of the sialic acid of the mitochondrial membrane with neuraminidase resulted in a considerable increase in the complement-fixing autoantigenic activity of the membrane without changing the apparent ability of the membrane to bind autoantibody in absorption experiments. 6. Exposure of mitochondrial membranes to autoantibody and complement caused an inhibition of both an inner mitochondrial membrane enzyme, i.e. cytochrome oxidase (48%) and an outer mitochondrial membrane enzyme, i.e. NADH cytochrome c reductase (rotenone insensitive) (37%).

Animals↗
PubMed1982

A comparison of the results of the brucellosis radioimmunoassay and other serological test in experimentally infected cattle.

The serology of 27 heifers found to be positive to culture after inoculation with Brucella abortus strain 544, was studied. Eighteen heifers had previously been vaccinated with strain 19 or strain 45/20 and nine were unvaccinated. Post-infection serum samples were tested for Brucella antibodies by radioimmunoassay (RIA), complement fixation test (CFT), indirect haemolysis test (IHLT) and Rose Bengal plate test (RBPT). All of the unvaccinated heifers showed strong humoral responses to experimental infection in the RIA, CFT, IHLT and RBPT. The CFT and RBPT became positive sooner after infection than the other tests in the unvaccinated heifers. However, in vaccinated heifers the RIA was the most sensitive test early in infection and the results of the RBPT were variable. Three of the vaccinated heifers showed weak and inconsistent humoral responses and, in these animals, the RIA gave fewer false negative reactions than the other tests.

Animals↗
PubMed1980

Clinico-hematologic and serological comparison of Japanese and Russian strains of Theileria sergenti.

The Japanese and Russian strains of Theileria sergenti were compared clinico-hematologically and serologically. They were subjected to passage in splenectomized calves 4 approximately 8 months old by the infectation of infected ticks or the inoculation of infected blood. As a result, the average of incubation of the Japanese strain was proved to be 23 days by the tick infection and 21 days by the inoculation of infected blood stored in a frozen state. The rate of detection and the average size of various forms of Theileria in the proliferative stage of the Japanese strain were as follows: Comma-shaped and bacilli forms, 86.8%; bacilli from 2.15 x 0.73 micron; oval and spherical forms, 11.3% and 1.97 x 1.22 and 1.46 micron, respectively; tetracoccal form and anaplasma-like form, 0.9 and 1.0%, respectively, and 0.54 micron in diameter. The highest parasitic rate of the Japanese strain was 11.9% on the average, and the rate of decrease in erythrocyte count 70.6% on the average. Rods were seen in protozoa particularly rich in cytoplasm. Such symptoms as pyrexia, depression, and anorexia appeared as Theileria propagated. No hemoglobinuria occurred or no schizonts were detected. Essentially the same results were obtained from the Russian strain as from the Japanese. Besides, Haemaphysalis longicornis produced in Japan acted also as a vector for the Russian strain. There was a good serological agreement between the Japanese and the russian strain in the results of the cross complement fixation test and the cross indirect fluorescent antibody test. There was no difference in the results of the complement fixation with the eight strains of the Japanese strain.

Animals↗
PubMed1977

Screening tests for syphilis. A comparison of the Treponema pallidum haemagglutination assay with two automated serological tests.

Two Treponema pallidum haemagglutination assay (TPHA) tests (Wellcome and Fuzizoki) were compared with the automated reagin test and the automated Reiter protein complement-fixation test in a routine screening system. The Wellcome TPHA was shown to lack sensitivity, and it is recommended that it should be used at a lower screening dilution. The Fuzizoki TPHA recorded a significant number of false positive and false negative results, and was therefore inferior to the fluorescent treponemal antibody absorption (FTA-ABS) test as a definitive test for syphilis. It is suggested that the TPHA be used in conjunction with a reagin test to replace some of the more inadequate screening tests currently in use. The automated screening tests were shown to have a similar sensitivity to the TPHA as well as several other advantages; they are recommended for testing large numbers of specimens.

Autoanalysis↗
PubMed1996

Characterization of a monoclonal antibody specific for Brucella smooth lipopolysaccharide and development of a competitive enzyme-linked immunosorbent assay to improve the serological diagnosis of brucellosis.

The reactivity of monoclonal antibody (MAb) 12G12 was analyzed in regard to the main biovars of Brucella species and some members of the families Enterobacteriaceae and Vibrionaceae which present serological cross-reactions with the smooth lipopolysaccharide (S-LPS) of Brucella species. This MAb was strictly directed against the common specific epitope of the Brucella S-LPS. It recognized all of the smooth Brucella strains and biovars except B. suis biovar 2. In order to improve the specificity of the serological diagnosis of brucellosis, a competitive enzyme-linked immunosorbent assay (cELISA) was developed with the horseradish peroxidase-conjugated MAbs 12G12 and S-LPS of B. melitensis Rev1. The specificity of the cELISA was analyzed with 936 serum samples from healthy cattle. The assay was evaluated with sera from heifers (n = 18) experimentally infected with B. abortus 544. After infection, the performance of the cELISA was in agreement with those of the complement fixation test and the rose Bengal plate test. Finally, the specificity of the assay was also evaluated in regard to false-positive serological reactions by using sera from heifers experimentally infected with Yersinia enterocolitica 0:9 (n = 4) and with field sera presenting false-positive reactions (n = 74). The specificity of the cELISA was greater than the specificities of the complement fixation test and the rose Bengal plate test. Indeed, the new assay detected only 31 of the 101 false-positive serum samples detected by at least one serological test.

Animals↗
PubMed1969

Properties of the Hong Kong influenza virus. I. General characteristics of the Hong Kong virus.

Biological properties of the Hong Kong influenza variants are described in relation to the isolation and identification of strains, sensitivity to nonspecific inhibitors, and serological diagnosis. Hong Kong virus strains were readily isolated in both eggs and monkey-kidney tissue cultures and were identified by haemagglutination-inhibition (HI) with chicken erythrocytes. The morphology, soluble antigen, and neuraminidase of Hong Kong isolates were similar to those of earlier A2 Asian influenza strains. Hong Kong haemagglutinins were related in varying degrees to previous human A2 and animal influenza viruses.The sensitivity of Hong Kong isolates to nonspecific haemagglutination inhibitors in serum varied widely. From least to most inhibitory, the ranking of sera of 8 animal species tested was: monkey, goat, chicken, human, rabbit, ferret, guinea-pig and horse. The same sera were treated with heat, trypsin, periodate, receptor-destroying enzyme and kaolin to determine the most effective way of removing nonspecific inhibitors. The results varied with the animal species involved.The A2 Asian antigen was nearly as effective as Hong Kong antigen in detecting antibody rises by HI tests. HI tests with either antigen were more efficient than complement-fixation tests for serodiagnosis of Hong Kong influenza, but for maximum efficiency both tests were required.

Animals↗
PubMed1970

The fixation of complement and the activated first component (C1) of complement by complexes formed between antibody and divalent hapten.

Hapten-antibody complexes prepared at equivalence with the bivalent hapten bis-DNP-octamethylene-diamine and purified rabbit anti-DNP antibody were fractionated by Sepharose gel-filtration and the fractions examined by electron microscopy. Individual fractions were tested for whole-complement fixation and C1 fixation. Dimer forms did not show this type of biological activity, while fractions containing tetramers and larger polymers exhibited both C and C1 fixation, which could be inhibited by prior exposure of the complexes to the univalent hapten epsilon-DNP-caproic acid. The dose-response result indicated that the C-fixation observed was not due to interpolymeric cooperative effects. It was concluded that in the generation of biological activity by soluble antigen-antibody complexes made with complement-fixing antibody, quaternary structural changes following specific combination with antigen may be as important as any tertiary structural alterations that occur in the individual immunoglobulin molecule.

Animals↗
PubMed1978

[Use of the passive hemagglutination test to diagnose toxoplasmosis].

The authors describe a method of obtaining toxoplasma erythrocytic diagnostic agent by sensitization of formalinized tannin-treated SRBC with purified toxoplasma antigen isolated by fractionation of complete toxoplasma antigen on Sephadex G-100. Comparative experiments with titration of sera of persons with suspected toxoplasmosis were conducted; the passive hemagglutination test with the antigen obtained proved to be highly sensitive in comparison with immunofluorescence and complement fixation tests.

Complement Fixation Tests↗
PubMed1988

Anticomplementary activity in serum of women with a history of recurrent pregnancy loss.

Viral complement fixation tests on women with a history of recurrent pregnancy loss were complicated by the presence of anticomplementary activity. This activity reflects the presence of a factor(s) in a patient's serum that nonspecifically fixes complement. When all patient sera tested were compared, 64.7% of women with recurrent pregnancy loss had anticomplementary activity compared with 22.0% among normal fertile pregnant women (p less than 0.01). In delineating when anticomplementary activity developed, it was found that 41.8% of women with recurrent pregnancy loss compared with 12.9% of normal pregnant women had this activity on entry to the study (p less than 0.01). This was primarily due to the fact that among women with recurrent pregnancy loss 50.0% of the pregnant versus 33.0% of the nonpregnant women had activity (NS). However, 55.2% of the anticomplementary negative women with recurrent pregnancy loss converted to a positive status compared with 15.4% of normal women (p less than 0.05). This was directly influenced by a conversion rate of 78.6% during pregnancy among women with recurrent pregnancy loss who entered the study nonpregnant and with no known cause for loss compared with a 33.3% conversion rate in their pregnant counterparts with recurrent pregnancy loss (p less than 0.025). Conversion to positive anticomplementary status occurred primarily by 20 weeks of gestation and appeared to be transient. Overall there was no association between the presence of anticomplementary activity and cervical colonization with genital mycoplasmas. The data suggest that women with a history of recurrent pregnancy loss develop a serum factor(s), usually by 20 weeks' gestation, that fixes complement. Thus these observations describe an additional anomaly in the immune system of women who experience recurrent pregnancy loss.

Abortion, Habitual↗