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PubMed1977

La Crosse virus soluble cell culture antigen.

A virus-free soluble antigen, obtained by ammonium sulfate precipitation of the supernatant fluids of La Crosse virus-infected BHK-21 cell cultures, was more reactive and more specific than infected suckling mouse brain antigen when compared by immunodiffusion and counterelectrophoresis tests. By complement fixation tests, the antigen was cross-reactive with heterologous California group arbovirus hyperimmune mouse ascitic fluids, but to a lesser degree than was the standard sucrose-acetone-extracted infected suckling mouse brain antigen. The major virion nucleocapsid protein of La Crosse virus was found by polyacrylamide gel electrophoresis to be the soluble antigen protein responsible for precipitation in immunodiffusion and counterelectrophoresis tests.

Antigens, Viral↗

PubMed1987

Comparison of serological tests for the detection of antibody to natural and experimental murine cytomegalovirus.

Three serological tests, i.e. complement fixation test, indirect immunofluorescent assay, and enzyme-linked immunosorbent assay (ELISA) were compared for sensitivity in the detection and titration of murine cytomegalovirus antibody. The three tests were compared using sera from experimentally inoculated and naturally infected mice bled at intervals from 3 to 140 days postinfection. In the acute infection, complement fixation and indirect immunofluorescent assay tests were of comparable sensitivity for early detection of antibody, whereas the ELISA was less sensitive. In persistent infection, higher titers were recorded with ELISA. Since murine cytomegalovirus has been shown to exert significant effects on the immune response of infected mice, this antigen should be included routinely in viral antibody screening programs.

Acute Disease↗

PubMed1997

[Comparison of the results from commercially available Brucella ELISA test kits for the investigation of bovine sera].

Bovine sera from a farm with bacteriologically confirmed brucellosis were taken for comparative serological studies using conventional methods (slow agglutination test--SAT, complement fixation test--CFT, Rose-Bengal-plate-agglutination-test--RBPT) and 5 different commercially available brucellosis ELISA testkits. These serum samples together with other infectious sera producing low level positive antibody titers in conventional tests did not react uniformly when tested by five different commercial ELISA testkits. The introduction of a standard control serum with a general cut-off resulted in a greater uniformity of ELISA results from 4 of 5 ELISA testkits and a closer approach to results of the CFT which is regarded as the confirmatory test for brucellosis. This control serum, on the other hand, was not suitable for the identification of the sensitivity of the various ELISA test kits. For this purpose, in SAT and CFT weakly positive reacting brucella sera from infected farms is needed.

Agglutination Tests↗

PubMed1992

Comparison of immunoblots with neutralizing and complement fixing antibodies in experimental and natural cases of visna-maedi.

In this study the humoral antibody response in visna-maedi virus disease in sheep during long-term infection was analyzed utilizing immunoblot assays, neutralization tests and complement fixation tests. In immunoblot assays antibodies to several virus specific protein bands were detected, both against the viral envelope glycoproteins and internal proteins of the virus. The immunoblot reaction pattern resembled that found in HIV-1 infection in humans, consistent with reported similar molecular weight of the major proteins of these two viruses. The immunoblot band pattern was compared with the pattern of complement fixing and neutralizing antibodies through the preclinical and clinical course in natural and experimental cases of visna-maedi. Of six immunoblot bands identified as virus specific, the antibody response against three gag products and the major env glycoprotein appeared early in infection, at a similar time as the complement fixing antibodies. The response against two proteins, one presumably the transmembrane protein and the other possibly a gag precursor, was delayed.

Animals↗

PubMedDate not supplied

[Development of the technology of the purification of Coxiella burnetii from components of culture medium].

The technology of the purification of C.burnetii from the yolk-sac components of the cultivation medium, including differential centrifugation, membrane filtration, enzymatic hydrolysis, the standardization of the preparation thus obtained and its stabilization by the method of lyophilization, was developed. Antigens, prepared in accordance with this technology, were sufficiently active and highly specific. The possibility of their use in different serological tests (the complement fixation test and the indirect fluorescent analysis) was studied. In addition, these antigens were found to have good capacity for sorption on the solid-phase carrier (plates) and to be suitable for use in such highly sensitive test as the enzyme immunoassay.

Animals↗

PubMedDate not supplied

[Passive hemagglutination test in the serodiagnosis of syphilis].

Passive haemagglutination test with pathogenic Treponema pallidum antigen in the serodiagnosis of syphilis, carried out on 752 sera (518 syphilitic patients and 234 subjects without evidence of syphilis) showed a greater general sensitivity as compared the cardiolipin tests and complement fixation test with Reiter protein antigen, and lower, to immunofluorescence (FTA--ABS). Its specificity is relatively lower than that of the cardiolopin and treponemal tests. Associated with the cardiolipin test it may be preferably used for the detection of syphilis, and moreover, passive haemagglutination may confirm the diagnosis in case of weak positive results with the FTA--ABS test. The latter is, however, essential for problem cases.

Antigens, Bacterial↗

PubMed1979

Antigenic variation among strains of the New Jersey serotype of vesicular stomatitis virus.

Four strains of vesicular stomatitis virus--New Jersey (Hazelhurst, Guatemala, Panama, and Concan) were compared by cross-neutralization and complement-fixation tests. They were indistinguishable by complement-fixation test; however, by plaque-reduction neutralization method, slight antigenic differences were observed between the Hazelhurst and the three other strains. It is concluded that these antigenic differences are insufficient to warrant reclassification of vesicular stomatitis virus--New Jersey into two distinct subtypes, as has been recently proposed.

Animals↗

PubMed1981

[Value of serodiagnosis in human trichinosis].

The paper with the comparative evaluation of serological tests in the diagnosis of human trichinellosis. The tests include ring precipitation test cold-precipitation test; agar diffusion technique; complement fixation test (cold binding); dermatol, carmine, and latex agglutination reactions. Serological tests applied to human trichinellosis were found to have different reliability. The most reliable reactions were those of ring precipitation, complement fixation test (cold binding) and cold- precipitation test. The agar diffusion technique was of little reliability only.

Agglutination Tests↗

PubMed1989

In vitro fixation of C3d and C5b-9 on platelets by human platelet reactive antibodies.

Most platelet-reactive autoantibodies and alloantibodies are not able to fix complement in vitro. However, exceptions have been found. These antibodies are usually characterized by the conventional platelet complement fixation test. A recently developed competitive enzyme immunoassay for quantitation of platelet-associated immunoglobulins and a modification thereof allowed the quantitative study of fixation of C3d and the membrane attack complex (C5b-9) on platelets by HLA antibodies, human platelet autoantibodies, and drug-dependent antibodies (ddab). The highest amounts of both complement products were fixed through ddabs, whereas autoantibodies only showed moderate complement fixation. This enzyme immunoassay is a valuable tool for the characterization of the complement-fixing properties of platelet-reactive antibodies.

Antibodies↗

PubMed1991

Anaplasmosis in Uganda. II. Prevalence of bovine anaplasmosis in Uganda.

The prevalence of bovine anaplasmosis was studied in 320 Zebu cattle randomly selected from three regions of Uganda: (Central, Southwestern and Northwestern) using dot-ELISA, Western immunoblotting, rapid card agglutination test (RCAT), capillary tube agglutination test (CAT), complement fixation test (CFT), and parasitological techniques. Dried blood on Whatman filter paper No. 1 was eluted in PBS 0.05% Tween 20 prior to testing at an initial dilution of 1:25. The prevalences of parasitaemia were 25% in the central region, 28% in the southwestern region, and 35% in the northwestern region, and the serological prevalence was lowest in the central region and highest in the northwest. Overall, prevalence rates obtained by dot-ELISA (61.9%) and Western immunoblotting (62.5%) were 1.5 times those obtained by RCAT (41%) and three times those obtained by CAT (22.5%). The overall prevalence rates obtained by dot-ELISA and Western immunoblotting compared favourably with the CFT data. The present data utilizing dried blood on filter papers indicate that there is a high prevalence of anaplasmosis in those regions of Uganda surveyed, and confirm our observations and those of others that collecting blood on filter papers is a suitable technique for large scale screening and for seroepidemiological studies.

Agglutination Tests↗

PubMed1999

Experimental infection of white-tailed deer with rangiferine brucellosis.

Experimental infections of white-tailed deer (Odocoileus virginianus) with Brucella suis biovar 4 were evaluated over a period of 6 wk. Five adult male hand-raised white-tailed deer were inoculated with 1 x 10(7) colony forming units of B. suis biovar 4 in the conjunctiva and serologically evaluated over 6 wk by the card test (CARD), rivanol test (RIV), serum agglutination test (SAT), complement fixation test (CFT), particle concentration fluorescence immunoassay (PCFIA), and competitive enzyme linked immunosorbent assay (cELISA), as routinely used for bovine samples. Six weeks postinoculation the animals were euthanized and cultured for B. suis biovar 4. One deer became serologically positive 4 wk postinoculation on CFT, CARD, PCFIA, and cELISA. At 6 wk postinoculation, CFT was positive in four infected deer, CARD was positive in three deer; RIV, SAT, and PCFIA was positive in two deer; and cELISA was positive in one deer. Only the CFT was 100% sensitive. At necropsy B. suis biovar 4 was isolated from four of five deer, and representative colonies were biologically similar to the challenge organism.

Agglutination Tests↗

PubMed1985

Characterization of a partially purified leptospiral genus-specific protein antigen.

A leptospiral genus-specific protein antigen (GP-Ag) was partially purified from Leptospira interrogans serovar kremastos strain Kyoto and canicola strain Hond Utrecht IV, by treating the organisms with Triton X-100 followed by a purification procedure including fractionation with DEAE-cellulose column chromatography and ethanol precipitation. Genus-specificity of the antigens from kremastos Kyoto and canicola was shown by the immunodiffusion (ID) test. Complement fixation (CF) test and enzyme-linked immunosorbent assay (ELISA) using GP-Ag from kremastos Kyoto showed the genus specificity of the GP-Ag. Sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) of the antigens from kremastos Kyoto and canicola showed a major band with Coomassie blue stain. The band of GP-Ag from kremastos Kyoto was also shown by immunoprecipitation using the monoclonal antibody (GP-7) to GP-Ag from kremastos Kyoto. The molecular weight of the genus-specific antigen was estimated to be approximately 62 000. The genus specificity of the antigen was reduced by heating at 100 degrees C or treatment with proteolytic enzymes, but not reduced by treatment with sodium metaperiodate.

Agglutination Tests↗

PubMed1975

Use of the complement fixation (CF) test in Lassa fever surveillance. Evidence for persistent CF antibodies.

A survey to detect individuals with antibodies to Lassa virus was undertaken among hospital personnel in the eastern and southern provinces of Sierra Leone late in 1974. Sera were evaluated by the complement fixation test. The data obtained showed that some contacts of Lassa fever patients in the 1972 epidemic had developed antibodies to the virus; individuals who had never reported being sick also showed evidence of infection, with significant CF antibody titres in their sera. Surviving Lassa fever patients from the 1972 epidemic still had easily measurable levels of persisting CF antibodies. The significance of these data is discussed; in addition it is recommended that the CF test should continue to be the method of choice in mass surveys for this virus disease until other tests can be developed.

Adolescent↗

PubMed1985

Reduction of non-specific reactions to the Brucella abortus serum agglutination test by the addition of EDTA.

Serum samples taken from cattle known to be infected with Brucella abortus, and samples routinely collected as part of the brucellosis eradication scheme were tested by the serum agglutination test (SAT), complement fixation test (CFT) and the SAT modified by the addition of ethylene diamine tetra-acetic acid (EDTA). In 64 per cent of the samples giving an SAT titre greater than 100 iu, but a CFT titre 8.3 icftu or less, the agglutination reaction was sufficiently affected by the action of EDTA for the titre to drop to below 100 iu. Only 5 per cent of samples giving an SAT titre greater than 100 iu and a CFT titre of 20 icftu or greater were affected in a similar manner by EDTA, and none of the 29 sera taken from known infected animals showed a drop in titre to below 100 iu, although some with titres greater than 500 iu did show significant EDTA sensitivity.

Agglutination Tests↗

PubMed1991

Observations on the eradication of Brucella ovis infection from a ram flock.

The measures taken to eradicate Brucella ovis infection from a naturally infected flock of 64 rams are described. Lesions of epididymitis were detected in 18 rams, all of which gave either positive or suspicious reactions in the complement fixation test. A further 20 rams gave serological reactions in the complement fixation test. Subsequently, semen was collected from 14 of these 20 rams and B. ovis was cultured from the semen of all 14 rams. Serum samples from two rams failed to react in the complement fixation test. However, they were identified as infected with the aid of an enzyme-linked immunosorbent assay and the subsequent culture of semen samples. It is suggested that, when eradicating B. ovis infection from ram flocks, the enzyme-linked immunosorbent assay be used in addition to both the complement fixation test and the physical examination. Using a combination of tests as described can increase the likehood of an earlier eradication of B. ovis infection.

Journal Article↗

PubMed1977

The effect of immaturity of the calf on immunological responses to strain 19 and killed 45/20 adjuvant vaccines.

Thirty brucellosis free calves with zero titres to the serum agglutination test (SAT), complement fixation test (CFT) and antiglobulin test (ABGT) were vaccinated with strain 19 at ages from seven hours to 198 days. Calves 75 days of age and older responded with normal serological patterns, developing high titres to all three tests. At 45 days and younger most calves responded with much reduced titres, some were negative to the SAT and CFT but all develped titres to the ABGT. Two of the younger group were subjected to an anamnestic test at about a year old and gave a positive response, indicating that the calf may be effectively primed with S19 as early as the first day of life. Three of the group were colostrumdeprived yet the patterns of their responses were similar to those of the colostrum-fed calves. Seventy-four zero titres calves were vaccinated with killed 45/20 adjuvant vaccine at ages from 60 to 320 days. Up to 200 days of age only seven of 33 calves gave positive response. From 200 to 280 days 18 of 29 responded and from 280 days of age all calves a positive response. The late development of competence to respond to this adjuvant vaccine is somewhat unusual and is discussed. It is suggested that the rough strain 45/20 may be a very weak antigen in cattle.

Age Factors↗

PubMed1972

Mycoplasma: growth precipitation as a serodiagnostic method.

A serological test is described which is designated the growth precipitation test and is based on the release of diffusible compounds from mycoplasma strains. During growth of mycoplasmas on solid substrates, precipitation lines are formed with antibodies from homologous antisera independent of heat-labile serum factors. The application of the test as a serodiagnostic tool was investigated by performing a complete serological cross-test with 10 mycoplasma strains. The test was compared with the growth inhibition test, the complement fixation test, and the indirect hemagglutination test. It is demonstrated that the growth precipitation test has a good degree of specificity. The species and subspecies used here showed cross-reactions between serologically closely related strains, but these heterologous reactions occurred only after longer incubation than the homologous reactions.

Acholeplasma laidlawii↗

PubMed1998

Suitability of various Coxiella burnetii antigen preparations for detection of serum antibodies by various tests.

Comparison of four serological tests (complement fixation (CF) test, microagglutination (MA) test, microimmunofluorescence (MIF) test, and enzyme-linked immunosorbent assay (ELISA)) for detection of post-infection antibody response in human and animal sera revealed a low sensitivity of the CF test with acute Q fever human, goat and sheep sera but not with chronic Q fever human sera and sera of aborting cows. The remaining three tests gave similar results with human (both acute and chronic) and cow sera, but the ELISA was more sensitive than the MA and MIF tests with goat and sheep sera. A treatment of phase I Coxiella burnetii (C.b.) cells with chloroform-methanol, potassium periodate and trichloroacetic acid (TCA), and mild acidic hydrolysis did not result in increase of the sensitivity of the tests when compared with the natural phase I and phase II C.b. cells, respectively. The suitability of various C.b. antigen preparations for the abovementioned serological tests with various sera is discussed.

Acute Disease↗