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Cytotoxic and agglutinating H-Y antibodies in multiparous female mice.

Pregnancy in inbred mice is associated with long-term accceptance of H-Y-incompatible skin grafts (in some "rejector" strain females) and with the formation of cytotoxic and agglutinating H-Y antibodies (in females generally). Yet there is no evident correlation between the two phenomena; postpartum females of nonrejector (H-2k), intermediate rejector (H-2d), and rejector (H-2b) strains all may produce H-Y antibodies, and postpartum H-2b females bearing H-Y-incompatible male skin grafts of long-standing may or may not produce H-Y antibodies depending on the individual recipient. Thus, it remains to be seen whether the presence of H-Y antibodies, as detectable to our sperm cytotoxicity and Protein A-SRBC agglutaination tests, signifies a corresponding presence of enhancing H-Y antibodies in pregnant females, and indeed whether enhancement (with respect to H-Y antigen) may play any part in the survival of the inbred male fetus.

Agglutination Tests↗

[Role of direct agglutination test (DAT) in the diagnosis of visceral leishmaniasis in Tunisia].

Direct agglutination test (DAT) is a simple ans economical technique for visceral leishmaniasis diagnosis. An evaluation, held in Tunisia with 41 confirmed visceral leishmaniasis cases and 58 negative controls, showed a sensitivity of 95.1% and a specificity of 96.5%. These results were highly correlated with those of ELISA and fluorescent antibody test. The use of DAT in our country will be very relevant, mainly in district hospitals, where others classical tests are not available. It will also be useful during sero-epidemiological studies on both human and canine visceral leishmaniasis.

Agglutination Tests↗

Relevance of direct agglutination test in seroepidemiological survey of Kala-azar in epidemic foci of Bihar.

The performance of direct agglutination test (DAT) was evaluated under field conditions in three endemic districts of Bihar. In three villages of these districts, 197 households were surveyed with a population of 1167 individuals. Out of 1167 individuals, 603 were clinically examined for VL and blood samples of each examined individual were collected for DAT. The proportion of DAT positivity in both males (32.33%) and females (32.94%) was statistically not significant (p > 0.05). The proportion of DAT positivity in all age groups was observed to be similar (p > 0.05). The GMRT was observed highest in active VL cases. Out of 20 DAT positive individuals having fever, 5 were examined parasitologically (bone marrow aspiration) and in three cases positivity was observed. This showed the relevance of DAT in early diagnosis of visceral leishmaniasis and usefulness of DAT in field conditions.

Adolescent↗

Antibodies to spermatozoa. VI. Comparative studies of sperm-agglutinating activity in groups of infertile and fertile women.

In order to evaluate the significance of sperm antibodies in relation to infertility, studies have been made on both infertile and fertile populations. The infertile population consisted of about 150 couples with long-standing infertility, predominantly of the unexplained type. The fertile population consisted of 78 antepartum women and 35 postpartum women. In all these individuals, sperm antibody was sought by two methods of serum testing, namely, the Kibrick (K-B-M) and the F-D methods of sperm agglutination. In the infertile group, the Kibrich test was positive for 23.1 per cent of the women and 9.6 per cent of the men, whereas the F-D test was positive in 16.4 per cent of the women and 5.0 per cent of the men. In contrast to these values, it was found in the fertile group that the Kibrick test was positive in 2.7 per cent of these women and the F-D test was also positive in 2.7 per cent of them. A number of technical details and precautions have also been emphasized. These principles are intended to minimize the occurence of nonspecific and false-positive results.

Agglutination Tests↗

Evaluation of microscopic agglutination test as a diagnostic tool during acute stage of leptospirosis in high & low endemic areas.

BACKGROUND & OBJECTIVES: Making a diagnosis on the results of a single microscopic agglutination test (MAT) is difficult because of the uncertainties about the cut-off titre. The present study was conducted to determine the significant titre for a single MAT in areas of high and low endemicity for leptospirosis. METHODS: A total of 1944 serum samples were collected from healthy individuals and confirmed patients residing in areas of high and low endemicity. All the sera were screened by MAT using 10 live leptospiral strains as antigens. From the distribution of titres among healthy individuals and in patients, the sensitivity and specificity at different cut-off titres were calculated. Likelihood ratio positive (LR+), likelihood ratio negative (LR-), and LR+/LR- were calculated. Receiver operating characteristics (ROC) curves were plotted for the early and late stages of the disease in both the areas. RESULTS: The ROC plot was totally below the no benefit line during the first week of illness in high endemic area. During the second to fourth weeks it showed better characteristics and the best cut-off titre was 1:200, where the sensitivity was 93.4 per cent and specificity 74.7 per cent LR+ LR- ratio was 41.82 indicating reasonable separation between the positive and negative test results. In the other states the ROC plot was above the no benefit line even during the first week, the best cut-off being 1:50 where the sensitivity was 56.7 per cent and specificity was 90.6 per cent. During the second to fourth weeks the test showed the best characteristics in the low endemicity regions with an ROC curve having the ideal shape. Best cut-off was at 1:100 where the sensitivity was 96.6 per cent and specificity 94.8 per cent LR+ LR- ratio was 523.25 indicating a wide separation between the positive and negative test results. INTERPRETATION & CONCLUSION: MAT does not have any diagnostic value during the first week, particularly in high endemic areas. The best cut-off to be used will be 1:50 in low endemicity areas during the first week, 1:100 during the second to fourth week and 1:200 in high endemicity regions during the second to fourth weeks.

Agglutination Tests↗

Anti-HIV-1 antibody testing using modified gelatin particle agglutination: a large field study.

Anti-HIV testing using gelatin particle agglutination (GPA) assay was investigated in parallel with ELISAs from routine service at Siriraj Hospital. In the first strategy, 174,032 sera from a patient population with an HIV-1 seroprevalence of 13.72 per cent were assayed using reduced volumes of GPA reagents, giving a cost reduction of 40 per cent. In the second strategy, 90,560 pregnant women and 48,936 emigrant workers with an HIV-1 seroprevalence of 2.2 per cent and 0.3 per cent, respectively, were tested in pools of 4 sera using the manufacturer's recommended volumes, giving a cost saving of 67 per cent. Overall, the sensitivity and specificity were almost identical with standard methods. Thus, parallel use of either modified GPA might be considered appropriate when testing large numbers of samples. However, both modified versions of GPA are not recommended as the first assay for diagnostic or blood bank screening especially in high prevalence of HIV infection.

Agglutination Tests↗

[A comparison of two tests for rheumatoid factor: latex test and l-agglutination (author's transl)].

Positive and negative reactions of latex test (LX) and streptococcal L-agglutination (LA) were correlated significantly with one another. However, in approximately 70% of the sera tested only one test was positive, LX at a much higher rate than LA. Postive rheumatoid factor tests were found in 117 patients with rheumatoid arthritis and in 595 patients with other diseases, exclusive of connective tissue diseases. Among the seropositive patients with rheumatoid arthritis, a positive reaction was seen in 85% and 71% by LX or LA respectively. Patients with nonrheumatic diseases showed positive reactions at a rate of 3,1%, with 2,3% and 1,7% in LX or La respectively. This shows that LA is less sensitive, but more specific as compared with LX. With increasing age of the patients, the rate of "nonspecific positive" reactors in patients with nonrheumatic diseases increases, more strongly for LX than for LA. The distribution of positive tests (LX positive, LA positive, or both positive) was analyzed in various groups of patients. The rate of positive reactions in both tests is significantly higher than patients with rheumatoid arthritis; it further increases with the severity of the disease. In cases with positive CRP, the rate of sera positive in both tests is higher, equally in patients with and without rheumatoid arthritis. In patients with nonrheumatic diseases an elevated serum gamma-globulin is associated with positive reactions in both tests and with positive LX. In all other grouping of patients, the rheumatoid factor tests did not yield significant deviations from chance distribution, as determined by chi2 analysis.

Age Factors↗

Establishment of immunoglobulin M(IgM)-immunosorbent agglutination assay (ISAGA) for diagnosis of toxoplasmosis.

AIM: To establish an immunosorbent agglutination assay (ISAGA) for detection of IgM antibodies against Toxoplasma gondii. METHODS: In the ISAGA, wells of microtiter plates were coated with anti-human IgM antibodies and sealed with 1% bovine serum albumin. After the test sera were added and incubated, the plates were washed, T. gondii tachyzoite antigen suspension was added, and incubated overnight at 37 degrees C. The ISAGA results were evaluated by comparing with those detected by Danish ISAGA and ELISA and those detected by slide enzyme immunoassay (S-EIA). RESULTS: Forty-four sera from Danish pregnant women were tested by the IgM-ISAGA, 41(93.2%) were consistent with the Danish results. Sixty-seven sera from Danish and Shanghai pregnant women were detected by IgM-IgM-ISAGA and S-EIA, the total consistency rate was 92.5%. A significant correlation was found between the titers of the ISAGA and S-EIA, with a Pearson correlation coefficient of 0.589(P < 0.001). The titers of ISAGA were eighteen times higher than those of S-EIA. This method enables the detection of IgM antibodies as low as approximately 0.08 IU/ml. CONCLUSION: The IgM-ISAGA is therefore sensitive, specific, easy to perform, and is useful for mass screening and diagnosing recent Toxoplasma infection or reactivation.

Agglutination Tests↗

[Standardization of neutralization tests using the COBL cell line and comparison with the particle agglutination test for measles serology].

The aim of the present study was the detection and comparison of measles antibody titers with particle agglutination (PA) and neutralization (Nt) methods, in the sera samples of 364 subjects from different age groups. PA method was performed with a commercial test kit (Serodiameasles, Fujirebio Com. Japan), and Nt test which was standardized in this study, by using COBL (cord blood) cell lines, has been started to use in our laboratory as a reference method. As a result, antibody titers detected by PA were in parallel to the titers which detected by Nt test, and it was concluded that the differences in antibody titers would arise from the differences of test principles and viral antigens.

Adolescent↗

[A new modification of the horse erythrocyte agglutination test for the detection of heterophilic antibodies in infectious mononucleosis].

The new modification (HEA test) is based on the presence of a singularly distinct agglutination of native horse erythrocytes in a serum dilution 1:10 and more from cases with Epstein-Barr virus infectious mononucleosis. In 1665 sera the HEA test was equally often positive as the OCH test (in 5.2%) whereas the IM test with formal-treated horse erythrocytes was positive only in 3.7%. The sensitivity of the IM test in comparison with the OCH test was 61%, the specificity 99.5%. The sensitivity of the HEA test in comparison with the OCH test was 76%, the specificity 98.7%. The specificity of the HEA test in a group of 52 sera without IgG antibodies to EB-viral capsid antigen (EB VCA) was 94%, in another group of 130 sera from hepatitis cases it was 98%. In 103 pairs of sera from infants aged 1-5 years the positivity of the OCH test, HEA test, IgM antibodies to EB VCA and the four-fold rise in the titre of EB-viral IgG were, 20%, 16%, 23% and 24% respectively. Analogous values in 159 pairs of sera from older subjects were 42%, 40%, 38% and 29%. The HEA test is recommended as useful adjunct to the OCH test for the detection of heterophile antibodies even in young children.

Agglutination Tests↗

[A carboxylated latex agglutination test for the serological diagnosis of human and animal brucellosis].

A carboxylated latex agglutination test was developed for detecting antibodies in sera of human and animals with Brucellosis. The latex was chemico-linked with B. melitensis 16M extracted with 5% NaCl. Comparing with physico-sorbent latex antigens in detecting antibodies in sera of human and animals with Brucellosis, the chemico-linked latex is more stable, more specific and more sensitive. In addition, preparation of chemico-linked latex and some factors affecting the result of the test were also described in the paper.

Agglutination Tests↗

Gelatin particle agglutination assay to detect anti-PGL-I antibodies in leprosy patients and in household contacts: a preliminary study.

A preliminary study of anti-phenolic glycolipid-I (PGL-I) IgM antibody detection using M. leprae gelatin particle agglutination (MLPA) test kit is described. Antibodies were demonstrated in 70% of our leprosy patients taking antileprosy treatment. The percentage of positivity of multibacillary cases was 86.0, whereas that of paucibacillary cases was 30.0. Good correlation was found between bacteriological index and the presence of antibodies. Antibodies were detected in 28% of our patients released from treatment. Fourteen out of 27 household contacts were found to have antibodies but none of the normal controls were seropositive. These preliminary data demonstrate that MLPA test is not applicable as sero-diagnostic test or as a test of cure, but may be useful for epidemiological studies and as a research tool.

Agglutination Tests↗

Detection of antibodies to human immunodeficiency virus (HIV-1) with the particle agglutination test.

This study was designed to evaluate the sensitivity and specificity of the particle agglutination test (PA) for antibody detection and to determine whether it is possible to use this test as a screening routine for anti-human immunodeficiency virus type I antibodies instead of the enzyme linked immunosorbent assay (ELISA). Serum samples were collected from 5,142 subjects and tested with the above two tests. A total of 83 samples were observed to be ELISA-positive. However, only 36 were found to be seropositive by the PA test. These samples were then confirmed by the Western blot analysis and 28 were positive for HIV-1, 33 were negative and 22 gave indeterminate results. The most common indeterminate band was p15/17 and the p24 band came next. The sensitivity and specificity of the PA test were 100% and 99.9%. The results indicate that the PA test is not only a convenient technique but also has a high sensitivity and specificity. It can be used as a primary screening test for antibody against HIV-1 instead of the ELISA.

Agglutination Tests↗

[The combination of mixed agglutination and immunofluorescence reactions for the simultaneous detection of 2 ABO-system antigens in isolated cells].

A combination of two immunologic tests, mixed agglutination and immunofluorescence, is suggested, that permits the detection of two AB0 system antigens in the same cell within one process. Such mode better validates the conclusion on the group appurtenance of cells, for instance, in analyses of mixed stains originating from different subjects with different blood groups.

ABO Blood-Group System↗

Validation of a simple and cheap gelatin particle agglutination test for human immunodeficiency virus using dried blood spot samples.

OBJECTIVES: To validate a Gelatin Particle Agglutination (GPA) test for HIV using dried blood spots with a view to applying the test in large epidemiological studies. DESIGN: A method comparison study with standard enzyme linked immunosorbent assay (ELISA) using the Recombigen HIV-1/2 kit as the gold standard. SETTING: Blair Research Laboratory, Harare, Zimbabwe. SUBJECTS: Sera and dried blood spots samples were available from 379 women from Mbare, Harare and Mupfure, Shamva District, Zimbabwe who had participated in HIV studies conducted by Blair Research Laboratory. MAIN OUTCOME MEASURES: Results of the GPA and Recombigen HIV-1/2 ELISA using serum and dried blood spots. RESULTS: With the Recombigen HIV-1/2 ELISA as the gold standard, sensitivity and specificity of the GPA were 100% and 99.2% respectively using serum. With dried blood spots sensitivity and specificity of the GPA test were 100%. The cost of analysing one sample, based on cost of reagents and accessory materials only, was Z$300 for GPA compared to Z$1,200 for ELISA. Furthermore, hands-on time was significantly reduced with the GPA compared to ELISA. CONCLUSION: The GPA method is simple, less labour intensive and much cheaper, yet is equally sensitive when compared to standard ELISA. The high sensitivity with blood spots makes the test ideal for large-scale epidemiological studies in remote rural areas with no infrastructure for advanced diagnostic methods.

Adolescent↗

Co-agglutination (Co-A): a rapid test for the diagnosis of experimental trichinellosis.

Co-agglutination test (Co-A test) which has a wide application spectrum in bacterial diseases has been used for the detection of some parasitic infections. This work was to evaluate Co-A test as a new diagnostic tool in detection of T. spiralis antigens in both sera and urine of experimentally infected mice as compared with ELISA, for the diagnosis of trichinellosis. The results obtained by Co-A test were closely correlated with those of ELISA. The most important was that both tests were capable to detect T. spiralis antigen in urine of light and heavily infected mice only at the 30th days post infection. So, Co-A is an accurate, easy and rapid test that can be used on large scales for diagnosis of trichinellosis.

Agglutination Tests↗

Direct agglutination test with urine samples for the diagnosis of visceral leishmaniasis.

A new direct agglutination test (DAT) for use with urine samples for the diagnosis of visceral leishmaniasis (VL) has been developed and compared with the conventional DAT with serum samples and our previously reported enzyme-linked immunosorbent assay (ELISA) with urine samples (urine ELISA). The new DAT, in which anti-human IgG was used as enhancing antibody, was tested with urine samples from 75 VL patients and 225 non-VL patients and healthy people. The sensitivity of the new DAT (90.7%), was almost the same as that of the conventional DAT (91.0%) and the urine ELISA (93.3%). The specificity of the new DAT (96.4%) was nearly identical with that of the urine ELISA (97.3%). A urine-based DAT has several advantages over the conventional DAT: sample collection is non-invasive and it can process larger numbers of samples with smaller amounts of antigen.

Adult↗