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PubMed1982

Complement-fixation test for rotavirus detection: comparison and analysis of different methods to reduce anti-complementary activity of some specimens.

The complement-fixation test may be used to detect rotaviral antigens directly in clinical specimens. However, a certain number of specimens tested for human rotaviruses by the complement-fixation test show an anti-complementary activity. By comparing eight techniques we analysed this anti-complementary activity and identified the best method for its reduction. Pretreatment of clarified supernatant of stool suspensions by some methods resulted in a reduction of anti-complementary activity, without reducing the sensitivity of the method. Clarified supernatants of 8/36 (22.2%) specimens were anti-complementary; this anti-complementary activity was best removed by absorption with fetal calf serum or calf albumin. Such treatment offers practical means of increasing the specificity of complement-fixation test. Some observations suggest that the anti-complementary activity of stool suspensions may be frequently due to presence of one or more chelating agents that may be in faecal specimens.

Absorption↗
PubMed1970

A new rapid method for automatic complement-fixation tests.

Several automatic methods have been described for complement-fixation tests. However, each of these methods presents some inconvenience either due to cross contamination or to limited sensitivity. The new method consists basically in separating clearly the fixation stage from the haemolytic stage by using two different sampler plates placed on the same sampler. By this procedure, it becomes possible to obtain a very high sensitivity. The serologist may set the temperature and the duration of the reaction according to the properties of the tests under study. In addition, the sampling rate can be raised to 120 analyses per hour.

Animals↗
PubMed1983

Comparison of the enzyme-linked immunosorbent assay and complement fixation test for detecting Brucella ovis antibodies in sheep.

A solid-phase, indirect, enzyme-linked immunosorbent assay (ELISA) was compared with the microtitre complement fixation test for detecting Brucella ovis antibodies in 220 ram sera. The ELISA was more sensitive than the complement fixation test; it demonstrated antibodies in 11 sera from known infected or vaccinated rams that were complement fixation test negative. No false positives were recorded with the ELISA and, in 36 sera positive to both tests, the ELISA titres were consistently higher than the corresponding complement fixation test titres.

Animals↗
PubMed1982

Confirmation of herpes simplex virus type 2 infections in herpes-like genital lesions by a simple complement-fixation test.

The presence of complement-fixing antibody to an early herpes simplex virus type 2 (HSV-2) antigen (the AG-4 antigen) was correlated with HSV-2 infection in the sera of patients with genital herpes. Eighty-eight per cent of sera taken two weeks after clinical diagnosis of a primary or recurrent herpes infection in patients, confirmed to have HSV-2 by virus isolation and typing, contained the anti-AG-4 complement-fixing antibody. None of the patients with genital HSV-1 had the antibody, and only 9% of controls or patients with facial HSV-1 infection had positive results for the antibody. This correlation was used to identify genital HSV-2 infections when either no virus sample had been taken or when virus isolations had been unsuccessful. Thus, a simple complement-fixation test can confirm an HSV-2 virus infection without isolation of the virus from the herpetic lesion.

Adolescent↗
PubMed1978

Studies on bovine leucosis V. A comparative study on the practical value of the agar gel immunodiffusion test, the indirect fluorescent antibody technique and the micro complement fixation test for the detection of antibodies to bovine leucosis virus.

The practical value of the indirect fluorescent antibody technique (I FAT), the micro complement fixation test (M CFT) and the agar gel immunodiffusion test (AGIDT) for the detection of antibodies to bovine leucosis virus (BLV) was investigated. For this purpose 1495 serum samples were examined. There was a remarkably good agreement between the three tests in the demonstration of antibodies to BLV especially if sera with a sufficient high concentration of antibodies ("late serum") were under investigation. These positive sera were derived only from farms which exploited one or more imported animals. A disagreement of results of the three tests was observed in 38 (2.5%) cases. This was due to: (1) difficulties in reading of the test; (2) presumably by the demonstration of different classes of antibody and (3) the fact that in the AGIDT sera could be used undiluted. This discrepancy was especially evident with sera with a low concentration of antibodies ("early or incubation sera"). A drawback for the M CFT is the anticomplementary activity found in 17% of the bovine serum samples.

Animals↗
PubMed1968

Enzootic pneumonia of pigs: complement-fixation tests for the detection of mycoplasma antibodies in the serum of immunized rabbits and infected swine.

The direct, the modified direct and the indirect complement-fixation tests were investigated as methods for the detection of antibodies for the enzootic pneumonia mycoplasma and for Mycoplasma hyorhinis in the serum of infected pigs and of immunized rabbits. Only the modified direct complement-fixation test in which the guinea-pig complement is supplemented with fresh, normal unheated calf serum was suitable for the detection of mycoplasma antibodies in sera of infected swine. Based on the close correlation between the production of typical lung lesions in experimentally infected pigs and the appearance of significant serum antibody titres, the modified direct complement-fixation test provides for the first time a sensitive, specific in vitro method for the detection of enzootic pneumonia in the live pig. This test also permitted the in vitro differentiation of the mycoplasma causing enzootic pneumonia from M. hyorhinis which causes polyserositis. Antibodies in the sera of rabbits were demonstrable by the ordinary direct complement-fixation test. However, in contast to the observation made with swine sera, only a slight quantitative antigenic difference between the enzootic pneumonia mycoplasma and M. hyorhinis was seen when the tests were performed with rabbit serum antibodiies.

Animals↗
PubMed1968

An evaluation of the Technicon Autoanalyzer for automating complement-fixation tests.

Obviously there is need for a reliable means of automating complement-fixation tests. This paper offers an account of experience with an AutoAnalyzer adapted for this purpose. Results of syphilis screening tests done by the AutoAnalyzer compared with those of tests performed manually showed satisfactory agreement, the automated method being slightly less specific in our hands. There were no false negative results with the AutoAnalyzer if both Maltaner and Reiter protein antigens were used. The current design of the AutoAnalyzer contains three major shortcomings if it is to be used for large numbers of complement-fixation tests. First and foremost is the relatively slow sampling rate compatible with adequate sample discrimination. Second is the necessity for frequently replenishing the machine with samples. Third is the need for adjusting or cleaning the flow-cell during use.

Autoanalysis↗
PubMed1979

Verification of tween-ether antigen for complement fixation test in the diagnostics of toxoplasmosis.

Tween-ether toxoplasma antigen for the complement fixation test was verified on a more extensive clinical material. From a series of 949 patient's sera, positive reaction was obtained in 44% of samples with the tween-ether antigen and in only 33.5% of samples with the FT antigen. All sera, giving positive results only with the tween-ether antigen, were also positive in the Sabin-Feldman test. The authors believe that by using this type of more sensitive antigen, containing also the cell-wall components of Toxoplasma gondii, it would be possible to standardize serological examination by the complement fixation test on the basis of an international standard serum.

Animals↗
PubMed1990

Evaluation of a modified complement fixation test and an indirect hemagglutination test for the serodiagnosis of melioidosis in pigs.

A complement fixation test modified by the addition of porcine serum and an indirect hemagglutination test were used to detect antibodies to Pseudomonas pseudomallei in pigs. These tests together with cultural examinations were carried out with 250 pigs. The sensitivity and specificity values were 79.3 and 99.5% and 82.8 and 93.2% for the modified complement fixation and hemagglutination tests, respectively. When results from the combination of both tests were considered, the values were 86.2 and 92.8%, respectively.

Animals↗
PubMed1981

Diagnosis of human rotavirus infections: comparison of an electrophoretic method, a modified complement fixation test and electron microscopy for rotavirus detection.

A modified complement fixation test and a counter-immuno-electrophoresis on cellulose acetate were compared to electron microscopy, in detecting Rotavirus from stool specimens. Out of 75 samples from patients with acute gastroenteritis, 40 per cent yielded positive results. The main advantages of the above two methods in routine diagnosis are discussed.

Complement Fixation Tests↗
PubMed1987

Effect of Eperythrozoon ovis on the lysis of sheep erythrocytes in the complement fixation test.

When erythrocytes from sheep experimentally infected with Eperythrozoon ovis were used in the titration of reagents for a standardised complement fixation test, increased amounts of both haemolysin and complement were required for erythrocyte lysis compared with preinfection titrations. The haemolysin requirement increased by up to 125% at 55 days post-infection and complement requirement increased by up to 40% at 40 days post-infection. These changes appeared to correlate with the development of a macrocytic anaemia in affected sheep rather than E. ovis parasitaemia. The results emphasise the need to carefully monitor the haematological parameters of sheep used as sources of erythrocytes for the complement fixation test.

Anaplasmataceae Infections↗
PubMed1984

A complement fixation test for visceral leishmaniasis using homologous parasite antigen I.

The complement fixation test using homologous parasite antigen was evaluated in 60 individuals with confirmed visceral leishmaniasis and compared with the results obtained in individuals with other parasitic and infectious diseases; 88% of the confirmed cases of visceral leishmaniasis were positive whilst no positive reactions were observed in individuals with other infectious and parasitic diseases. The specificity was partially compromised by anticomplementary activity in 5%, limited to individuals with visceral leishmaniasis, and doubtful (2%) or sero-negative (7%) results.

Antigens, Protozoan↗
PubMed1978

A complement fixation test for antibody to the contagious equine metritis organism.

A complement fixation test (CFT) based on that used for brucellosis (Brinley Morgan and others 1971) has been developed for use on the sera of horses exposed to the contagious equine metritis (CEM) organism. None of 50 single samples from horses thought to be unexposed to the CEM organism was positive to the test, although five showed inconclusive reactions. Samples were examined from 41 mares either proved to be infected or from an infected stud. Of these 21 were positive, 11 were inconclusive and nine were negative. The relationship of the CFT to reactions in the other tests used in this condition is discussed--the serum agglutination test (SAT) and antiglobulin test (AGT) (Benson and others 1978). Complement fixing antibodies appear to remain for a longer period in the infected animal, and are therefore more likely to be of use in the diagnosis of the chronic carrier state.

Agglutination Tests↗
PubMed1986

A short, reliable, highly reproducible complement fixation test for the serological diagnosis of contagious equine metritis.

A complement fixation test, using round-bottomed microtitration plates and an 8 channel microdiluter, based on that used for brucellosis by Herr, Huchzermeyer, Te Brugge, Williamson, Roos & Schiele, 1985, has been developed for use on the sera of horses to detect antibodies to the contagious equine metritis organism. The results with 2 known positive sera tested 116 times in 27 separate tests were reproducible for the most part within a twofold range. They seldom exceeded these limits and never exceeded a fourfold range. The test itself is capable of being carried out within 90 min. The test was slightly more sensitive when sera were inactivated in a hot air oven for 50 min at 58 degrees C, as compared to inactivation at 62 degrees C in a water-bath for 50 min. There were no false negative or false positive reactions and no anticomplementary activity in the sera tested.

Animals↗
PubMed1983

Improvement of complement fixation test antigen for the diagnosis of Mycoplasma hyopneumoniae infection.

Several attempts were made to develop a complement fixation test antigen which is useful and practicable for the diagnosis of mycoplasmal pneumonia of swine. For the preparation of antigen, Mycoplasma hyopneumoniae was grown in broth medium containing horse serum for 14 days by shaking culture. Anticomplementary activity of antigen was eliminated by addition of complement and heating at 56 degrees C for 30 minutes. Storage of this antigen at 4 degrees C or at room temperature for 12 months had no significant effect on antigen titer. In the complement fixation test with this antigen, complement-fixing antibodies could be detected up to 41 weeks after experimental inoculation by KIO4 treatment of serum. They were demonstrated in 81.4% of 296 sera collected at slaughterhouses in five prefectures.

Animals↗
PubMed1991

A new complement fixation test for the diagnosis of neurocysticercosis in cerebrospinal fluid.

As a modification of the classical complement fixation test, a new test for immunodiagnosis in cerebrospinal fluid (CSF) was developed. When the assay was used for the diagnosis of neurocysticercosis in 149 CSF samples from patients and 1036 from controls, results showed 93% concordance with positives and 97% with negatives when compared with the results obtained by ELISA; in addition, it was positive in 12 of 16 cases of neurocysticercosis in which negative results had been obtained by ELISA. The new complement fixation test is particularly useful in CSF because of the immunological peculiarities of the subarachnoid space, where local synthesis of oligoclonal antibodies is induced by infectious agents. Reagents used for the assay are easy to produce and preserve, and the test is inexpensive, reliable and easy to perform. As this test detects the immunobiological consequence of the antigen-antibody reaction, it can be used in conjunction with other assays, such as ELISA, that measure the reaction directly, thus increasing the diagnostic possibilities. It may also be used where financial or technical limitations hinder access to other immunodiagnostic tests.

Animals↗