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The nature of the antibody response to Yersinia enterocolitica serotype IX in cattle.

The nature of the antibody response of cattle to the antigen of Yersinia enterocolitica IX cross-reacting with Brucella spp. was examined. Density-gradient ultracentrifugation, ion-exchange chromatography, antibody adsorption and elution and disulphide bond reduction tests showed that both 19 S IgM and 7 S IgG(1) and IgG(2) antibodies were produced in response to the cross-reacting antigen. The highest titres of cross-reacting antibodies were detected by the agglutination and Coombs antiglobulin tests. Production of complement-fixing and precipitating antibodies cross-reacting with Br. abortus was transient and high titres were not attained.In contrast, although infection with Br. abortus also evoked cross-reacting antibodies of the IgM and IgG classes, much higher titres were produced in the complement fixation and precipitation tests and these persisted for long periods. At all stages of the serological response to both organisms, the two infections could be differentiated by the quantitative Rose Bengal plate test.

Animals↗

Evaluation of A newly developed DOT-ELISA kit for the diagnosis of human brucellosis.

A new dot-ELISA kit for detection of Brucella antibodies in human sera was developed and compared with that of serum agglutination test, Rose Bengal plate test, rapid slide agglutination and Coomb's antiglobulin test. Following testing of 120 human sera from suspected patients of occupational risk, 25 gave positive reaction in Rose Bengal plate test, 25 in rapid slide agglutination test, 26 in serum agglutination test, 27 in Coomb's antiglobulin test and 28 in dot-ELISA kit. Dot-ELISA kit picked up more positive than any other Serological test, indicating its superiority over the other laboratory tests for the diagnosis of brucellosis.

Agglutination Tests↗

Long-term follow-up of indirect hyperbilirubinemia in full-term Turkish infants.

OBJECTIVE: This retrospective follow-up study was performed to evaluate the suitability of the recently reported exchange transfusion limits (serum indirect bilirubin level of 428-496 mumol/1, 25-29 mg/dl) for Turkey. MATERIAL AND METHODS: The study groups totalled 102 children, 8-13 years of age, who had been born at term with birthweights greater than 3000 g and had been treated for indirect hyperbilirubinemia during their newborn period; the control group consisted of 27 children of the same age-group without indirect hyperbilirubinemia. Children were grouped according to their maximum serum and bilirubin levels and direct Coomb's test results. Physical and neurological examinations, visual and brainstem auditory evoked potentials and the Wechsler Intelligence Scale for children--Revised for Turkish Children were performed. RESULTS: There was no difference between the groups with regard to mean visual and brainstem auditory evoked potential latencies. Children whose direct Coomb's tests were positive had significantly lower IQ scores and more prominent neurological abnormalities (p < 0.05). IQ scores and prominent neurological abnormalities did not differ among the other groups. Nine children had prominent neurological abnormalities associated with abnormal brainstem auditory evoked potentials. An important risk factor was the duration that the infant's serum indirect bilirubin level remained greater than 342 mumol/l (20 mg/dl). CONCLUSION: The current limit of 342 mumol/l should continue to be used for infants whose direct Coomb's tests are positive in our country. Until better criteria for exchange transfusion other than the indirect bilirubin level are established, the current limits should also still be followed for infants whose direct Coomb's tests are negative in Turkey, where regular neonatal follow-up examinations are not satisfactory.

Adolescent↗

[Coombs-positive reactions in mice during transplantation of syngeneic tumors].

The effect of the growth of syngeneic transplantable tumors on Coombs' positive test in mice was studied. The modification of the hemagglutination test permitting one to minimize the amount of reagents used is described. Tumor transplantation induced Coombs' positive reactions in most recipients. The phenomenon could be observed in 4 murine strains and 6 tumor systems. Sometimes autoimmune reactions occurred before the emergence of palpable tumors. It is concluded that despite the influence of nonspecific factors, the principal cause of autoimmune reactivity is tumor growth. It seems that the appearance of alien "normal" histocompatibility antigens is a characteristic feature of all the tumors; besides, the host response pattern to these antigen is cross-reactive, including autoimmune component. Coombs' positive test may be one of numerous manifestations of such an autoimmune process.

Animals↗

Circulating antisperm antibodies in recurrently aborting women.

One hundred seventy-three women with a history of three or more recurrent consecutive abortions were analyzed for circulating antisperm antibodies with a radiolabeled antiglobulin assay (RAA), a modified enzyme-linked immunosorbent assay (ELISA), a tray agglutination test (TAT), and a sperm immobilization test (SIT). No pregnancies were subsequently gestated to term in women who were antisperm antibody-positive unless they were inoculated with their husband's leukocytes as treatment for an immune basis (not related to antisperm antibodies) for their recurrent abortions. In women with an immune basis for their recurrent abortions, immunization with leukocytes from their male partners increased the ability of these women previously aborting their fetuses to carry their fetuses to term, even if they had positive results in the ELISA, TAT, and SIT; women with positive results in the RAA continued to abort subsequent pregnancies, despite leukocyte immunization. Immunization of antisperm antibody-positive women with their partner's leukocytes did not incite or increase the antisperm antibody titer, with any of the assay techniques.

Abortion, Spontaneous↗

Comparison of mixed antiglobulin reaction and direct immunobead test for detection of sperm-bound antibodies in subfertile males.

OBJECTIVE: To compare two methods of detection of surface bound antibodies on spermatozoa from subfertile males. DESIGN: Prospective comparison of direct mixed antiglobulin reaction (MAR) for immunoglobulin (Ig)G with direct immunobead test (IgG and IgA) applied to spermatozoa from male partners of infertile couples. Circulating unbound antibody measured by tray agglutination test in serum and seminal plasma in a representative proportion. SETTING: Seminology laboratory. PATIENTS: One hundred nine male partners of infertile couples. RESULTS: Highly significant correlation between direct MAR (IgG) and direct immunobed test (IgG) and between both of these tests and serum unbound antibody measured by tray agglutination test. Highly significant correlation between direct immunobead test (IgA) and seminal plasma unbound antibody measured by tray agglutination test, but no correlation with MAR (IgG). CONCLUSIONS: Mixed antiglobulin reaction (IgG) is a cheap, quick, and sensitive screening test, but immunobead test (IgA) provides useful additional information on class of antibody on spermatozoa that may be clinically more important.

Agglutination Tests↗

The indirect mixed antiglobulin reaction test using a commercially available kit for the detection of antisperm antibodies in serum.

A simple procedure is described for the detection of antisperm antibodies of the IgG class in human serum using the indirect mixed antiglobulin reaction (MAR) test. The test uses only one dilution of serum (1/16) and no washing procedures. The test was performed in 361 sera. Comparative studies were conducted with the tray agglutination test (TAT) in 164 sera and with the direct MAR test on semen in 110 cases. In 24 sera with borderline test results, and in 13 sera with discordant findings, evidence of antisperm antibody activity was investigated by means of additional testing, namely, the adenosine triphosphate release cytotoxicity test (ARCT) and the indirect immunobead tests (IB) for IgA and IgM. Using the value of 40% reaction between motile spermatozoa and coated latex particles as the lower limit of significant activity, the indirect SpermMAR test has a sensitivity of 96% and specificity of 87%. The two false-negative indirect MAR results occurred in cases with IgM, the clinical significance of which is doubtful. Hence, the indirect MAR test appears to be an ideal screening procedure for sperm antibodies in serum since it is easy to perform and sensitive. Considering the false-positive rate of 13%, additional tests should, however, be performed on sera with a reaction of 40% or more in order to assess the relevance of the antibodies as a cause of infertility.

Agglutination Tests↗