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PubMed1981

In vitro responses of lymphocytes from cattle with advanced Mycobacterium paratuberculosis infection to homologous and heterologous antigens.

The accuracy with which the purified lymphocyte and whole blood modifications of the lymphocyte stimulation test were able to detect animals with clinical Johne's disease was compared with that of the complement fixation test and microscopic faecal examination. Confirmatory diagnosis of Johne's disease was based upon histopathological examination of intestinal tissue. False positive results were obtained only with the complement fixation test. The only test not giving rise to false negative results was the purified lymphocyte modification of the lymphocyte stimulation test using johnin at 20 micrograms/ml and avian tuberculin at 20 micrograms/ml or 2 micrograms/ml. The whole blood technique was less accurate than the purified lymphocyte technique for the in vitro detection of cell-mediated immune responses to antigens. The purified lymphocyte technique appears to have potential as an diagnostic test for Johne's disease in cattle and merits further evaluation.

Animals↗
PubMed1988

Response of heart rate to atropine and left ventricular function in Chagas' heart disease.

Chagasic patients may have a normal or abnormal response of heart rate to atropine. To determine if this response to atropine is related to the degree of left ventricular dysfunction, we studied 33 patients with a positive complement fixation test for Chagas' disease. Eleven subjects, with atypical chest pain and negative complement fixation test, were used as controls (sero-negative). Left ventricular wall motion and the left ventricular volumes were determined by ventricular cineangiography. Coronary arteriography was also performed. The sero-negative subjects had normal left ventricular wall motion and the left ventricular diastolic volume was 88 +/- 24 ml/m2. The response of heart rate to atropine was 50 +/- 8 (mean +/- SD) (range 40-65 beats/min). On the basis of the response to atropine, the chagasic patients were divided into groups with a normal response (greater than 40 beats/min) and those with an abnormal response (less than 40 beats/min). Sixty five per cent of those with a normal response had left ventricular apical aneurysms. The left ventricular end-diastolic volume was not significantly different from the sero-negative subjects (96 +/- 26 ml/m2). Six patients (30%) had a left ventricular diastolic volume between 110 and 140 ml/m2. Sixty two per cent of those chagasic subjects with an abnormal response had diffuse left ventricular hypokinesis, and the left ventricular end-diastolic volume was 192 +/- 49 ml/m2 (P less than 0.01). The response of heart rate and the left ventricular diastolic volume were inversely correlated in the chagasic patients (r = -0.88, P less than 0.01). Our results indicate that myocardial damage and the degree of left ventricular dilatation are more severe in chagasic patients with an abnormal response of their heart rate to atropine. Furthermore, the inverse correlation between these two variables is highly indicative of a relationship between the response of heart rate to atropine and the degree of left ventricular dysfunction.

Adult↗
PubMed1975

[Serological studies on rabbits inoculated with Toxoplasma gondii]

Serological studies on toxoplasmosis were conducted with rabbits sera that were immunized with RH strain or infected with Beverley strain of Toxoplasma gondii. Complement fixation tests, agar-gel double diffusion tests and agar-gel immunoelectrophoresis were performed. Toxoplasma crude antigen was prepared from the organisms in mice peritoneal fluids, which were infected with RH strain of Toxoplasma gondii. The organisms were suspended in saline volume originally exudated and counted in hemocytometer for purity of the organisms over 99%. These suspended organisms were prepared by sonication, and the solution was centrifuged for 30 min. at 10,000 rpm in 4 degrees C. These supernatant fluids were used as crude antigen. On the other hand, purified antigens were fractionated on Sephadex G-200 gel filtration. A Sephadex G-200 column, 80 by 4 cm, equilibrated with Tris-HCl-(0.1M)-NaCl (1.0M) buffer, pH 8.0 was used. The eluate fractions were collected in 3 ml per hour and the absorbance at 280 nm was measured with a Beckman Du-2 spectrophotometer. Each tube is pooled into 6 fractions by protein density graph. For immunization of rabbits, crude antigen of RH strain was emulsified with an equal amount of incomplete Freund's adjuvant and l ml of mixture was injected subcutaneously into them once a week for 5 successive weeks. Antisera were obtained at an interval of a week, beginning the first week after the last immunization, while several rabbits were infected with Beverley strain of Toxoplasma gondii by inoculating about 200 cysts and antisera were obtained from them serially at a week interval. The results were as follows: 1. The sera from the rabbits immunized with the RH strain or infected with Beverley strain of Toxoplasma gondii againist the crude antigen showed the first positive reactions in 2 or 3 weeks after the administration or immunization in complement fixation tests. Maximum titers appeared in 4 or 5 weeks after immunization with RH strain and in 7 or 9 weeks after infection with Beverley strain respectively. 2. Complement fixation tests showed the positive reactions in the rabbits sera immunized with RH strain against the purified antigens II, III, IV, V and VI: moreover, antigen IV fraction showed the highest titer. On the other hand in the rabbits sera infected with Beverley strain against the purifed antigens II, III and IV fractions showed the positive reaction; especially, antigen fraction IV showed the highest titer. 3. In immuno-diffusion tests, the sera from the rabbits immunized with RH strain and infected with Beverley strain, against the crude antigen appeared the precipitin bands 2 weeks after the immunization or infection. And the former showed the 2 or 5 precipitin bands after 5-8 weeks and the latter showed the l or 2 precipitin bands after 6 weeks. 4. The sera from the rabbits immunized with RH strain against the purified antigens II, III, IV,V and VI showed the precipitin bands, and the sera from the rabbits infected with Beverley strain against the purified antigens II, III and IV showed the precipitin bands in the immuno-diffusion tests. Especially antigen IV was the strongest reaction against the sera from RH strain and Beverley strain. 5. In agar-gel immunoelectrophoresis, the immunized sera against the crude antigen showed 8 arcs. But the infected sera against the crude antigen showed 4 or 5 arcs. 6. The immunized sera against the fractionated antigens II, III, IV, V, VI showed arcs, but against the fractionated antigen IV showed 6 arcs and in the antigens II, III, V, VI showed l or 2 arcs only. On the other hand, the infected sera against the fractionated antigens IV showed 4 arcs, II and III showed the l arcs, which was the most weak of all.

Journal Article↗
PubMed1988

Epidemiologic studies on schistosomiasis japonica in a newly found habitat of Oncomelania snails in Japan.

Epidemiologic studies' on schistosomiasis japonica were carried out in the Obitsu River basin of central Boso Peninsula, a newly found habitat of Oncomelania snails in Japan. Forty-five (4.1%) of 1,102 schoolchildren and 64 (10.4%) of 618 junior high school students were positive for the skin test with veronal buffered saline-extracted Schistosoma japonicum antigen. None was positive by the enzyme-linked immunosorbent assay, complement fixation test, double diffusion test in agar, and circumoval precipitin test. About one fifth of 356 adult inhabitants showed positive reactions in the enzyme-linked immunosorbent assay and complement fixation test as well as in the skin test, although only 25 individuals (7%) were positive for the circumoval precipitin test. All of 70 individuals who had shown positive reactions in at least two of the immunological tests were negative for a series of 5 stool examinations with the AMS III method. Release of S.japonicum cercariae was not observed in 15,000 O. hupensis nosophora collected in this area. Repeated examinations by dipping mice into irrigation ditches were negative. These observations, with the fact that all of 15 patients reported from nearby hospitals to have schistosome eggs in their tissues were over 45 years old, suggest that schistosomiasis has become a disease of the past in this area.

Adolescent↗
PubMed1979

Paratuberculosis in cattle and free-living exotic deer.

Paratuberculosis was studied among dairy cows and exotic deer that shared grazing areas at Point Reyes National Seashore, California. Of the 10 dairy herds tested, 5 (50%) were infected with Mycobacterium paratuberculosis (based on results of fecal culture). Mycobacterium paratuberculosis was cultured from 9 (8.7%) of the 103 bovine fecal samples and from 4 (3.9%) of the 103 bovine rectal mucosa scapings tested. Of 89 fecal samples from 52 axis deer (Axis axis) and 37 fallow deer (Dama dama), 5 (9.6%) and 3 (8.1%), respectively, contained M paratuberculosis. Culture of intestinal necropsy samples from the same deer indicated that 3 (5.8%) of the axis deer and 2 (5.4%) of the fallow deer were infected with M paratuberculosis. The cows were tested for serum antibodies by the complement-fixation test and by radioimmunoassay. Of 95 sera tested by complement fixation, 15 (15.8%) were positive, as were 15 (14.7%) of 102 sera tested by radioimmunoassay. Culture results and serologic test results were compared on a herd basis.

Animals↗
PubMedDate not supplied

[Comparison between fixation test with BCG and immunofluorescence in the diagnosis of leishmaniasis].

The complement fixation test with an antigen from commercially lyophilized BCG and the immunofluorescence test were carried out on serum samples from patients with visceral leishmaniasis, oriental sore and tbc. The IFT positivity was obtained only in patients with visceral leishmaniasis. The CFT with BCG showed positive results in 3 on 5 serum samples from patients with visceral leishmaniasis, in 2 with oriental sore, at low titres, and in 5 serum samples from patients with tbc. The AA. confirm the high specificity of the IFT, but suggest that the CFT with BCG can be used as a convenient screening test.

Antibody Formation↗
PubMed1980

[Comparative action of the low-molecular fractions from thymus and cerebral cortex polypeptides on the immune response in mice].

Rabbit antisera against low-molecular weight polypeptides from the thymus (thymosin and thymarin), brain cortex (cortexin) and white matter of the calf brain were cross absorbed with these polypeptides and tested in the complement fixation test with the above preparations. In addition they were also tested in the complement-dependent cytotoxicity test with thymocytes and bone marrow cells. Thymosin, thymarin and cortexin were shown to be antigenously similar but to differ in antigenous structure from the polypeptide of brain white matter. The biological effect of the polypeptides from the thymus and brain cortex is related to the thymus-dependent lymphocyte population rather than to B cells. Cross absorption has revealed that antisera against the polypeptides from the thymus and brain cortex contain antyibody both against common antigens and antigens specific only for the appropriate preparation. The antigenous set of thymarin responds better to thymic antigens than cortexin.

Animals↗
PubMed1973

Field evaluation of the gel diffusion test for the diagnosis of ram epididymitis caused by Brucella ovis.

A comparison was made with the microslide gel diffusion technique, the complement fixation test, cultural isolation, and clinical examination in detecting ram epididymitis caused by Brucella ovis. The results of the gel diffusion method are shown to be similar to those obtained by the complement fixation test and isolation of B. ovis from cultures of semen. The technique offers a reliable diagnostic method adaptable to field use in controlling ovine brucellosis and is more practical.

Animals↗
PubMed1975

Cellular immunity and circulating antibody to herpes simplex virus in subjects with recurrent herpex simplex lesions and controls as measured by the mixed leukocyte migration inhibition test and complement fixation.

Cellular immune responses and circulating antibody levels to herpes simplex virus were examined in patients with recurrent herpes simplex (HSV) infections and controls. Mixed leukocyte migration inhibition by herpes simplex antigen was less in affected patients than in controls but serum antibody levels were higher. There was no significant difference in leukocyte migration between patients with active or recent lesions and other herpes subjects, and the response in the mixed leukocyte migration test to phytohaemagglutinin (PHA) was similar in patients and controls. The data presented suggest that a localized defect in cell mediated immunity to herpes simplex virus may exist and be responsible for recurrent herpes simplex infections.

Antibodies, Viral↗
PubMed1976

[Diagnosis of human hydatidosis by hemagglutination and complement fixation (Weinberg) tests].

397 sera were tested by passive hemaglutination and weinberg reactions to diagnose hydatid cysts. 5 of 211 sera (2.53%) gave positive and 206 sera (97.63%) gave negative results by passive hemagglutination test, whereas in weinberg reaction 204 of them (96.68%) were negative and 7 (3.32%) anticomplementary. 13 sera (14.13%) from another batch of 92 sera gave negative results and 79 of them (85.87%) remained negative by passive hemagglutination test, when 6 of them (6.52%) were positive and 96 (93.48%) negative by weinberg reaction. 56 (91.80%) of 61 sera taken from patients operated on for hydatid cysts gave positive and 5 (8.20%) of them negative by passive hemagglutination test. 29 (47.54%) were positive, and 32 (52.46%) negative by weinberg reaction. In another batch of 15 sera taken from patients, 3 were positive and 12 were negative by passive hemagglutination, 2 were positive and 13 were negative by weinberg test. When the two tests are compared, passive hemagglutination test gave better results both qualitatively and quantitatively than weinberg so that it may be preferred in laboratories having improved facilities.

Complement Fixation Tests↗
PubMed1974

Value of the concomitant use of complement fixation and immunodiffusion tests in the diagnosis of coccidioidomycosis.

Complement fixation titers of 1:2 and 1:4 with coccidioidin are not always of diagnostic significance. The concurrent use of the complement fixation and immunodiffusion tests is an effective means for specific serologic diagnosis of coccidioidomycosis in patients with low levels of complement-fixing antibodies. In routine testing, it is recommended that only sera that are negative for complement-fixing antibodies at 1:8 but positive in the immunodiffusion test with coccidioidin be selected for titration at the 1:2 and 1:4 levels.

Antibodies, Fungal↗
PubMed1981

Notes to the standardization of complement fixation reaction with Toxoplasma antigen.

Optimum conditions are described for the complement fixation tests, including the method of primary incubation in a refrigerator, use of serial four-fold dilutions for complement titration and serial double dilutions for haemolysin titration. An optimum haemolytic system, most sensitive to the complement and 50% unit of complement is to be found at the same time. More than 100 sera were tested simultaneously using other reactions for toxoplasmosis (microprecipitation in gel, indirect fluorescence reaction and latex test) in order to verify the appropriate dose of complement for complement fixation reaction. It was also investigated to what extent small deviations in erythrocyte concentration in the haemolytic system may influence the results of serum titration. The appropriate dose of complement was found to be five 50% units. It is recommended to correct colorimetrically the concentration of erythrocytes in the haemolytic system. The questions of an appropriate serum dilution in the titration are discussed and a rapid and exact method is chosen. A simple method of reading 50% haemolysis in complement fixation microreaction is described.

Animals↗
PubMed1980

[Application of the serological clostridium assay in tumour diagnostics (author's transl)].

The suitability for serologic tumour diagnostics of the non-oncolysing strain Clostridium butyricum CNRZ 528 has been investigated. By introducing the transplantable Brown Pearce tumour of the rabbit and spontaneous tumours of the dog as test models the antibody production against clostridial rods--indicating malignant growth as had been proved with the Möse strain Cl. oncolyticum M55--has been evaluated. It could be established--utilizing the complement fixation test--that one serological method alone is not sufficient for obtaining clear-cut test results in anyone case. It is to be recommended that the indirect hemagglutination and the complement fixation tests ought to be combined. Our studies on specific clostridial antigens and on optimization of techniques will be continued.

Animals↗
PubMed1990

[Frequency of adenovirus group complement-specific antibodies in children].

Two hundred and six serum samples collected from children in Hacettepe University and Marmara University, Faculty of Medicine Hospitals, aged between 0-15 were tested for the presence of Adenovirus group specific antibodies by complement fixation test. The frequency of Adenovirus group specific complement fixation antibodies were found to be 4.85%. Most of positive sera were detected in 0-2 age group.

Adenovirus Infections, Human↗
PubMed1974

Detection of a protein of avian leukoviruses in uninfected chick cells by radioimmunoassay.

A radioimmunoassay (RIA) has been used to measure the quantity of protein P27 (mol wt 27,000; group specific-1 [gs-1]) of avian leukoviruses in different types of chicken embryos and tissues of adult chickens. The RIA used was 200- to 300-fold more sensitive than the complement fixation test and was able to detect as little as 0.3 ng of P27. Among six embryos tested, which are negative for gs antigen by the complement fixation test, P27 was undetectable in three embryos, but another three contained about 5 ng of P27 per mg of cell protein by RIA. The amount of P27 in gs antigen-positive cells ranged from 22 to 57 ng per mg of cell protein. P27 was found in liver, lung, ovary, feather pulp, and spleen from adult gs antigen-positive chickens. This protein was undetectable in various tissues from gs antigen-negative chickens.

Journal Article↗
PubMed1986

An evaluation of diagnostic methods for bovine brucellosis in Libya.

A total of 8,607 animals were examined for detection of brucellosis using blood serum agglutination as well as complement fixation tests. out of 8,607 sera samples examined, 125 sera played positive serum agglutination reactions in significant levels for brucella (1.45%). The percentage of reactors to the complement fixation test in the examined cattle was 1.82%. The relation between positive or doubtful cases of serum tube agglutination as regards to their complement fixation results were discussed in detail.

Agglutination Tests↗
PubMed1979

[Echinococcosis.--Current problems of diagnosis and epidemiology (author's transl)].

The indirect hemagglutination-test (IHA) and the indirect immunefluorescence-test are two modern methods for the serological diagnosis of an echinococcosis. If both tests are employed, the disease can be diagnosed with great reliability. A titer of 1 : 128 in the IHA makes the diagnosis of an echinococcosis very likely; a titer exceeding 1 : 1000 is significant. Serological tests of 36 patients with E. cysticus and E. alveolaris are evaluated; the interpretation of the results is discussed. In all patients the diagnosis was established between October 1977 and October 1978. In Germany the complement fixation test is commonly used to diagnose an echinococcosis. This test, however, is unreliable and fails in one third of all cases. It should be--as well as the Casoni skin-test--replaced by more sensitive and specific reactions. If the complement fixation test is positive, it might be used for postoperative controls. Compared to previous years the disease-rate among Germans is rising. The main reason seems to be the increasing tourism to foreign countries. Serological tests in small laboratories are unreliable and make an evaluation difficult. The diagnosis should be established in specialized centres.

Complement Fixation Tests↗