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Field utility of phenolic glycolipid coated latex agglutination test for rapid detection of bacilliferous leprosy cases.

Serum samples were collected from eighty-three leprosy patients and twenty-five healthy controls supposedly not exposed to Mycobacterium leprae infection. Phenolic glycolipid-1 coated latex agglutination test (PGL-LAT) was carried out with the serum samples to detect antibodies specific to M. leprae. Samples showing positive agglutination were 50% in the lepromatous leprosy (LL) group showing no erythema nodosum leprosum (ENL) complications, 66.6% in LL group with ENL complication, 60% in borderline lepromatous (BL) group, 50% in borderline (BB) and 33.3% in borderline tuberculoid (BT). The patients belonging to the tuberculoid (TT) group and most of the long-term treated patients were interestingly negative, and so were sera from all the healthy controls. PGL-LAT developed by us therefore is specific and a fairly sensitive technique to detect antibodies specific to M. leprae and will be very useful in field conditions.

Agglutination Tests↗

Correlation studies on Widal agglutination reaction and diagnosis of typhoid fever.

A total of 80 patients at the University of Uyo Teaching Hospital (UUTH) suspected of having enteric infections were screened for the presence of Salmonella species using blood, urine and stool samples along with Widal agglutination tests. Although 39 patients tested positive for the Widal agglutination test with titers ranging from 1:80 to 1:320, no Salmonella organism was encountered in some cultures. Statistical analysis revealed significant differences (chi2) at the 5% probability level between the Widal test and the cultures of the clinical samples. The results suggest that serological investigations alone may not be a reliable index for the diagnosis of Salmonella infections.

Adolescent↗

Evaluation of a rapid slide agglutination test for the diagnosis of acute canine leptospirosis.

The standard method for the diagnostic of leptospirosis is the microscopic agglutination test (MAT). Nevertheless, it is time-consuming and presents risk of infection for laboratory personal. Several attempts to simplify and reduce the time, skill and expense required for serological testing were made. Rapid slide agglutination tests (RSAT) have been used nowadays as a reliable screening test for the detection of acute and recent infections by many laboratories. The purpose of that study was to evaluate a new antigen preparation for the rapid diagnoses of acute canine leptospirosis and to compare it with MAT results in clinically suspect serum samples. Two hundred and thirteen serum samples from dogs of both sexes and different ages were tested and 141 (66.2%) animals were considered as positive by MAT. The most frequent serovar was icterohaemorrhagiae, followed by canicola. RSAT results were very similar to those observed on MAT, with 139 (65.3%) reactive animals. Correlation between RSAT and MAT was positive (0.82) and significant (p < 0.01). Concordance of results was of 93.4% (199/ 213) of the animals correctly diagnosed by RSAT. Considering MAT as the standard test, sensitivity of RSAT was calculated on 94.3% (133/141) and specificity on 91.7% (66/72).

Agglutination Tests↗

Comparison of three commercial rapid agglutination test kits for identification of coagulase positive staphylococci from foods and animals.

Three rapid agglutination assays for the identification of Staphylococcus aureus Monostaph (Bionor A/S, Skien, Norway), Staphyslide-Test (BioMerieux, Lyon, France) and Staph-Rapid-Test (Roche, Basel, Switzerland), were compared. A total of 104 Gram-positive, catalase positive cocci were tested: Nineteen Staphylococcus reference strains comprising 15 spp. (4 strains were coagulase positive), and 7 Micrococcus reference strains comprising 4 spp.; 22 food isolates comprising 13 S. aureus, 8 coagulase positive Staphylococcus spp., and 1 Micrococcus sp.; 56 animal isolates comprising 11 S. aureus, 9 S. hyicus subsp. hyicus, 2 S. intermedius, 15 coagulase positive and 19 coagulase negative Staphylococcus spp. Totally 54 strains were coagulase positive. Considering agglutination of a coagulase positive strain as a correct identification, Monostaph, Staph-Rapid-Test, and Staphyslide-Test correctly identified 52 (96.3%), 47 (87.0%) and 48 (89.0%) of the coagulase positive staphylococci, respectively. Monostaph, Staph-Rapid-Test and Staphyslide-Test showed 1 (2.0%), 4 (8.0%) and 4 (8.0%) false positive reactions respectively. Monostaph, Staph-Rapid-Test and Staphyslide-Test gave 0 (0.0%), 6 (5.8%) and 7 (6.7%) non-interpretable reactions, respectively. Monostaph may be a good alternative to the tube-coagulase test for rapid and reliable identification of coagulase positive staphylococci from both food and veterinary sources. However, false negative reactions may occur with coagulase positive strains of S. hyicus subsp. hyicus and S. intermedius.

Agglutination Tests↗

Anti-HIV-1: extended validation of a modified gelatin particle agglutination assay, and its use with an image analyser.

Modification of a commercial gelatin particle agglutination assay for anti-HIV-1 (MAT) has increased sensitivity, reduced reaction time and lowered the cost by 90%. We tested over 10,000 blood donations in parallel with the ELISA currently in use, and found the modified test to be highly sensitive and to enhance cost-effectiveness as it substantially reduces false-positive rates. The patterns of agglutination are clearly reproducible and readable by the naked eye and/or Image Analyzer, which provides objectivity and hard-copy documentation of results.

Agglutination Tests↗

[A method for the passive agglutination of polymer dispersions for the diagnosis of legionellosis].

The method of synthetizing dispersions of gelatin-modified polyacrolein microspheres 1.5-2.5 micrograms in diameter, used as a solid-phase carrier for the preparation of immunodispersion diagnostica, has been developed. The possibility of using immunodispersion diagnosticum in the passive agglutination test with hyperimmune rabbit sera has been demonstrated. High activity and specificity of immunodispersion diagnosticum, combining methodological simplicity and rapidity characteristic of agglutination, has been shown.

Agglutination Tests↗

Differentiation of newly isolated strains of Trypanosoma (Schizotrypanum) cruzi by agglutination and precipitation reactions.

Serological relationships between new isolates of Trypanosoma cruzi from a single locality have been examined by direct agglutination and immunodiffusion, and compared with established strains. The strains were divided into three groups according to the absorption properties of their precipitinogens. The three groups were defined by the same characteristics as those in which strains of wide-ranging provenance and long laboratory history have been placed by previous workers. A lack of correspondence between the antigenic constitutions of the groups as described here and as described previously was demonstrated using reference strains. Relatedness values were calculated from agglutination titres, which indicated a complex heterogeneity of antigens, and from which a scheme of relationships was drawn.

Agglutination↗

Cold-induced granulocyte agglutination. A cause of pseudoleukopenia.

Transient cold agglutination of her granulocytes developed in a 60-year-old woman with a left upper lobe pneumonia during the acute phase of her illness. This phenomenon was manifested by pseudogranulocytopenia, multiple clumps of granulocytes on her peripheral blood smear, and abnormal distribution of granulocytes and monocytes on the white blood cell histogram when measured on an automated hematology analyzer (Coulter S-Plus IV, Coulter Electronics Inc, Hialeah, Fla). The cause is postulated to be an IgM autoantibody directed against components of the granulocyte membranes. Spurious leukopenia is encountered infrequently with automated hematology analyzers. Cold-induced granulocyte agglutination should be recognized as a potential cause of pseudogranulocytopenia so that white blood cell counts can be accurately reported and unnecessary evaluation of patients for leukopenia can be avoided.

Agglutination↗

[Comparative study of antitoxoplasmic IgG isotypes titrated by high sensitivity direct agglutination and indirect immunofluorescence: consequence of the choice of methods on the expression of results in international units].

The use of International Units per ml (IU/ml) to express antitoxoplasmic IgG antibody titers in the various diagnostic systems presently proposed, is misleading owing to discrepancies in the values found from one test to the other for a given serum. The authors compared the results of high sensitivity direct agglutination (HSDA) to those of indirect immunofluorescence (IIF) in two studies, systematic and longitudinal, dealing with 158 sera stratified for values ranging from 102,400 to 5 IU/ml. Discordances between the methods, which are greater for high values, prompt the use of low-titer sera for standardization. From the systematic study, a correlation table was established and proposed to convert the HSDA results into the theoretical corrected values close to those that would be obtained by IIF. Although this may be of interest in maintaining a coherent language, this table has its limits, particularly in acute episodes where the various antibody kinetics vary and amplify further the discrepancies. In such situations, it seems advisable for both clinicians and biologists to raise any equivocal kept going by what is termed as International Units. Consequently, if the titers obtained by one method cannot be correlated to those of the technique of reference (IIF, Dye test), on the basis of using I.U., it would be appropriate to express the results in units related to the method or kit used (e.g. U/ml/HSDA for high sensitivity direct agglutination). Finally, whatever the technique, it is still mandatory to conserve a significant threshold value of protective immunization, common and identical to those classically adopted (8-12 IU/ml).

Adult↗

Elimination of mycoplasmal plate agglutination cross-reactions in sera from chickens inoculated with infectious bursal disease viruses.

Sera from chickens inoculated with various challenge infectious bursal disease viruses or infectious bursal disease vaccines were found to cross-react in the Mycoplasma gallisepticum (MG) and Mycoplasma synoviae (MS) serum plate agglutination (SPA) tests. Two-fold dilutions of these cross-reacting sera with phosphate-buffered saline before retesting eliminated all non-specific agglutination in the MG and MS SPA tests. Cross-reactions were observed in the SPA test using sera from chickens inoculated with either MG or MS. Dilutions of these sera 1:2 had little effect on the number of these cross-reactions. At 1:4 serum dilutions, however, the number of cross-reactions between MG and MS was reduced. At 1:8 dilution of test sera, cross-reactions between MG and MS were further reduced. Some reduction in specific MG and MS SPA reactions, however, also occurred at the 1:8 dilution of sera with some of the plate antigen.

Agglutination Tests↗

Immunofluorescence, enzyme-linked immunosorbent assay, particle agglutination and western blot for the detection of antibody to human immunodeficiency virus type 1.

Immunofluorescence assay (IFA) has been applied for detection of antibody to human immunodeficiency virus type 1 (HIV-1). To compare the IFA with an enzyme-linked immunosorbent assay (ELISA) and particle agglutination (PA), we examined the antibody response to HIV-1 in 475 sera from AIDS, PGL and ARC patients as well as several risk groups and healthy persons by three methods. The positive results by any methods were confirmed by western blot (WB). The results by all methods were well correlated on the sera from 45 asymptomatic male homosexuals and 70 female prostitutes. There were some false positive results by ELISA in the sera from prisoners and healthy persons. Four sera from drug abusers were positive only by PA and IFA and were negative by ELISA. All were WB-inconclusive. Particle agglutination and IFA results were compared with western blot analysis on 208 ELISA-positive sera. All IFA-strongly positive sera (84%) were positive by western blot. The sera with weakly positive, negative and inconclusive results by IFA (16%) were possibly any of positive, inconclusive or negative by western blot. By PA, 200 of 208 (97%) sera were PA-positive and 1% of these sera were WB-inconclusive while the PA-negative sera were either negative or inconclusive by western blot. These results suggested that PA is a simple and sensitive method for screening of HIV-1 antibody while IFA could be a primary confirmatory test and western blot would then be used for confirming any IFA-negative or inconclusive results.

Agglutination Tests↗

Increased agglutinability and haemadsorption of Marek's disease HPRS line 1 lymphoblastoic cells after concanavalin A treatment.

The HPRS line 1 lymphoblastoid cells derived from an ovarian lymphoma induced by Marek's disease virus in chickens displayed increased agglutinability and haemadsorption when incubated with Con A. The increase was proportional to the concentration of Con A used. The same Con A-untreated cells and normal chicken lymphocytes incubated with Con A exhibited no increase in agglutinability and haemadsorption, however.

Agglutination↗

[Lectin agglutination for the characterization of intestinal treponema isolates from different animal species].

The study presents the results of lectin-associated agglutinations of intestinal strains of treponemes isolated from pigs, dogs, mice and rats. At all 95 isolates are investigated, within the type strains for Treponema hyodysenteriae serotype 1-4 and Treponema innocens. Reactions with Concanavalin A and the Limulus-polyphemus-Lectin are often seen (twenty lectins used). Three out of four type strains for Treponema hyodysenteriae show an identical pattern of reaction, which is often seen in Treponema strains from dysentery suspected cases, too. Basing on these data a ranging in groups of lectin agglutination is proposed.

Agglutination Tests↗

Lectin mediated agglutination of Candida kefyr cells and their spheroplasts grown in sterol enriched growth medium and its correlation with lipid composition.

Supplementation of the growth medium with erosterol, cholesterol and lanosterol enriched the Candida kefyr cells, presumably cell membranes with sterols. Sterol enriched C. kefyr cells showed a decrease in percentage of PHA and Con-A mediated agglutination. Sterol supplementation also increased the sterol: phospholipid ratio and in such cells unsaturated fatty acids predominated over saturated ones. The overall effect of these changes resulted in rigidifying the cell membranes as indicated by shift of break in Arrhenius plots of Mg2+ ATPase. This showed that lectin mediated agglutination of yeast cells may be affected by its membrane fluidity.

Agglutination↗

Characterization of sperm agglutinating monoclonal antibody and purification of the human sperm antigen.

A monoclonal antibody capable of inducing tail-to-tail agglutination of human sperm was purified by ammonium sulfate fractionation and chromatography on DEAE-cellulose. A 75- to 100-fold purification was achieved. The antibody was of the IgG class and composed of gamma 1 heavy chain and lambda light chains. The purified preparation was effective in inducing sperm agglutination at a minimum concentration of 0.55 microgram/mL and was active after storage at 4 degrees C for over 3 months in the presence of sodium azide. The sperm antigen interacting with this YWK-1 antibody was purified by immunoaffinity chromatography on Sepharose 4B conjugated with YWK-1 antibodies. Two interacting proteins were isolated, with estimated Mr of 84 and 20 kD. The 84 kD protein had an isoelectric point in the range of pH 5.7 to 6.1, and probably lacks carbohydrate moieties. The 20 and 84 kD components may share common antigenic determinants.

Antibodies, Monoclonal↗

Lipid composition and lectin-mediated agglutination of Candida kefyr cells grown in media enriched with choline or its analogues.

Candida Kefyr cells and their spheroplasts grown in media enriched with choline, N,N'-dimethylethanolamine (DMEA) or ethanolamine (EA) showed decreased concanavalin A (Con A) and ph to hemagglutinin (PHA) mediated agglutination while supplementation with N-monomethylethanolamine (MMEA) increased PHA-mediated agglutination. In all cases, the amount of phospholipid was increased and, consequently, a decrease in the free sterol; phospholipid ratio was observed except in the case of MMEA where this ratio remained almost unchanged. In the cells grown in media enriched with choline. DMEA or MMEA, but not in the EA-supplemented medium, the phosphatidylcholine to phosphatidylethanolamine ratio was increased. Saturation of fatty acids as well as their chain length decreased, which could lead to increased membrane fluidity. No breakpoint in Arrhenius plots of Mg2+ -ATPase of the choline-supplmented cells was observed. Cells grown in media enriched with EA and MMEA did not show any sharp break in Arrhenius plots of Mg2+ -ATPase.

Agglutination↗

[Analysis of the result of thyroid microsomal particle agglutination test by ELISA].

No accurate method to detect thyroid microsomal (MC) antibody (Ab) in serum has been generalized. In this study, the titer of MC Ab obtained by the method of MC autoantibody particle agglutination (MCPA) was analyzed by enzyme linked immunosorbent assay (ELISA). MC and thyroglobulin (Tg) were prepared from Graves' thyroid. ELISA was done by coating the plate with MC, adding Tg to buffer and using peroxidase-conjugated anti-h IgG. 1) The titer of MCPA correlated with the MC Ab ELISA index in serum without Tg Ab, but it did not in serum with Tg Ab. MC Ab was negative by ELISA while it was positive by MCPA in some of the sera with Tg Ab. 2) When ELISA was done using buffer without Tg, the amount of IgG bound to MC was greater in serum with TGPA: + and MCPA: - than in serum with MCPA: + and TGPA: -. 3) The zone phenomenon observed in MCPA did not always indicate an excess of MC Ab. 4) MC Ab was positive by ELISA in some of the negative MCPA sera obtained from patients with Hashimoto's disease in which diagnosis was confirmed by biopsy. In conclusion, the result obtained in MCPA now in use is strongly influenced by Tg Ab. Furthermore, since binding of Ab to MC is judged by agglutination of particles in MCPA, ELISA is superior in sensitivity and accuracy in detecting MC Ab.

Agglutination Tests↗

Study of the plate agglutination test with rose bengal antigen for the diagnosis of human brucellosis.

This study presents the results of a number of serological tests, in particular the plate agglutination test using acid antigen stained with rose bengal, in the examination of 440 human sera for brucellosis. In addition, the sera were examined for the presence of specific antibodies of various classes (IgM and IgG). These investigations showed the rose bengal plate agglutination test to be highly specific and sensitive as a rapid method for the diagnosis of human brucellosis.

Agglutination Tests↗