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Lectin-mediated agglutination of murine lymphoma cells. Cell surface deformability and reversibility of agglutination by saccharides.

Agglutination of S49 mouse lymphoma cells by Ricinus communis I agglutinin can be reversed by the competing haptenic saccharide, lactose, soon after agglutination, but after further incubation in the absence of lectin the agglutination reaction could not be reversed by lactose and the cells remained as multicell aggregates. The irreversibility of S49 cell agglutination was time, temperature and lectin concentration dependent and its onset correlated with ultrastructurally observed deformation of adjacent cell surfaces and an increase in the proportion of adjacent cell surface areas in close apposition within multicell aggregates. Pretreatment of S49 cells with cytochalasin B or cytochalasin B plus vinblastine enhanced R. communis I agglutinin-mediated agglutination, while vinblastine alone and fluoride plus azide had essentially no effect. When drug-treated cells were agglutinated and then incubated in lectin-free drug-containing media for various times prior to lactose addition, the drug effects were more pronounced. Cytochalasin B alone or with vinblastine inhibited lactose reversal of S49 cell agglutination compared to the drug-free controls, while fluoride plus azide enhanced hapten reversibility. Electron microscopic analysis revealed that the onset of agglutination irreversibility correlated with cell surface deformation in the drug-treated cells. Cell aggregates that were more readily reversible by lactose (fluoride plus azide) were unchanged or less deformed, while S49 aggregates treated with cytochalasin B plus vinblastine were more deformed compared to controls without drugs. These experiments suggest a role for cell surface deformability as an important secondary effect during lectin-mediated cell agglutination of S49 lymphoma cells.

Agglutination

Mating reaction in Saccharomyces cerevisiae. VII. Effect of proteolytic enzymes on sexual agglutinability and isolation of crude sex-specific substances responsible for sexual agglutination.

The effect of proteolytic enzymes on sexual agglutinability of haploid cells of the yeast Saccharomyces cerevisiae was examined. Sexual agglutinability of cells of both a and alpha types was lost on treatment with alkaline protease and two kinds of neutral proteases of Bacillus subtilis, pronase and alpha-chymotrypsin. Agglutinability of alpha type cells was lost after treatment with acid protease of Rhizopus chinensis and trypsin, but that of a type cells was not. These results indicate that the sex-specific substance responsible for the sexual agglutination (agglutination factor) in a type cells differ from that in alpha type cells. Agglutination factors were solubilized from cell-wall fractions of both mating types by Glusalase treatment. These crude factors specifically inhibited the agglutinability of cells of the opposite mating type with little effect on the agglutinability of cells of the same mating type.

Agglutination

Latex agglutination, counterimmunoelectrophoresis, and protein A co-agglutination in diagnosis of bacterial meningitis.

Specimens of cerebrospinal fluid from 201 patients with meningitis caused by Neisseria meningitidis groups A/B/C/135, Streptococcus pneumoniae (23 types), and Haemophilus influenzae type b were tested for the presence of specific bacterial antigens by latex agglutination, counterimmunoelectrophoresis (CIE), and protein A co-agglutination. Specific antigens were found in 75% of the specimens by latex agglutination and CIE, and in 60% of the specimens by protein A co-agglutination. Non-specific reactions in protein A co-agglutination were prevented by heating the specimens to 100 degrees C for a few minutes. The three methods are simple and quick to perform. The smallest amount of antiserum was used in protein A co-agglutination, but we found this method less sensitive. Latex agglutination and CIE proved to be equally sensitive, but the first method was easier to adopt in practice.

Agglutination Tests

A quantitative assay for concanavalin A- and Ricinus communis agglutinin-mediated agglutinations of rat ascites hepatoma cells. Relationship between concanavalin A binding and cell agglutination.

A simple quantitative assay method was developed for the agglutination of rat ascites hepatoma cells mediated by Concanavalin A or Ricinus communis agglutinin. This method was based on the principle that the turbidity of a cell suspension is proportional to the sum of the cross-sectional area of cells and aggregatesmas predicted by the theoretical consideration, the turbidity decreased when cells were aggregated and the decrease was a function of the average number of the cells in aggregates. The agglutinability of the cells, judged by this method, showed a maximum value at a certain concentration of the agglutinin. By further addition of the agglutinin, the agglutinability slightly decreased from the maximum. These phenomena were observed both for Concanavalin A and Ricinus communis agglutinin. The binding and the agglutination experiments using [3-H]concanavalin A revealed that the binding to approx;0% of the total receptors caused a maximal agglutination. This suggested that the receptors responsible for the agglutination constitute only a small part of the total receptors on the surface.

Agglutination Tests

Methods for serotyping nasopharyngeal isolates of Haemophilus influenzae: slide agglutination, Quellung reaction, countercurrent immunoelectrophoresis, latex agglutination, and antiserum agar.

Nasopharyngeal isolates of H. influenzae were typed by the slide agglutination test, the Quelling reaction, the latex agglutination test, countercurrent immunoelectrophoresis, and the antiserum agar test. These tests gave essentially comparable results, with countercurrent immunoelectrophoresis and latex agglutination being slightly more sensitive. Cross-reactive problems encountered with latex agglutination and the expense of performing countercurrent immunoelectrophoresis or the antiserum agar test made these tests less practical than the slide agglutination test to identify single strains that were already isolated. The Quellung reaction and slide agglutination were the most rapid tests used to type an organism. For mass screening of multiple samples, countercurrent immunoelectrophoresis was the simplest technique. The antiserum agar test was slow but was the best technique to screen nasopharyngeal swab cultures to identify the presence of any encapsulated strains in the mixed flora. Whether any of the above techniques were as sensitive as the immunofluorescence test was not evaluated in this study.

Agar

An analysis of Con A-mediated agglutination in a Chinese hamster ovary subclone which responds morphologically to growth in dibutyryl cyclic AMP. III. The role of microvilli in the agglutination process.

We have used the H-7w subclone of a Chinese hamster ovary cell line (K1) to investigate the role of cell surface architecture (specifically microvilli, blebs, and sheets) in determining the relative agglutinability of a cell line with Con A. Our evidence clearly demonstrates that no specific, immediately recognizable surface architecture is associated with the agglutinable or non-agglutinable phenotype. Our data suggest that the expression of microvilli on the cell surface is neither necessary to nor sufficient for the phenotype described by enhanced agglutinability with Con A. Furthermore our work demonstrates that cells covered with blebs are as agglutinable as cells covered with microvilli thereby suggesting that the intertwining of microvilli may not be an essential facet of the agglutination phenomenon.

Agglutination

Ox erythrocyte agglutinability. 4. The effect of neuraminidase treatment on the agglutinability of cells and ghosts.

1. The inherited differential agglutinability of cattle erythrocytes is shown to be similarly expressed on ghosts and intact cells. 2. Removal of virtually all sialic acid by prolonged neuraminidase treatment does not alter the agglutinability status of ghosts prepared from either high or low agglutinable cells. Hence the differing sialic acid content of the two cell types is not responsible for the differential agglutinability. 3. The significance of these findings with respect to other well defined agglutination systems and current theories of membrane structure is discussed.

Animals

Comparison of the tray agglutination technique with the gelatin agglutination technique for the detection of spermagglutinating activity in human sera.

Extensive comparisons were made between the tray agglutination test (TAT) and the gelatin agglutination test (GAT) in order to evaluate the validity of the TAT for demonstrating spermagglutinating activity in human sera. The incidence of spermagglutination in both tests was about the same. In screening 118 samples from infertile men and women, the TAT seemed to be more sensitive, whereas the reverse was true when 143 samples earlier found positive with the GAT were tested. These differences were mainly encountered with low titers. Two T-agglutinating sera (or 2% of the sera) with a GAT titer of 16 or more (both had a titer of 64) were missed with the TAT. On the other hand, two sera (or 2% of the screened samples) having small H-agglutinates up to a titer of 32 to 64 were positive only with the TAT.

Agglutination Tests

The agglutination reactions of Haemophilus paraphrophilus and H. paraphrohaemolyticus, and some observations on the agglutination of H. Aphrophilus and H. haemoglobinophilus (H. canis).

Agglutination tests were used to study the surface antigens of two recently described species of the genus Haemophilus, H. paraphrophilus and H. paraphrohaemolyticus, and their antigenic relationship to other members of the genus. The results obtained with a few strains of H. haemoglobinophilus and H. aphrophilus are also reported. The species H. paraphrophilus appears to be homogeneous; no major cross-reactions were observed. The species H. paraphrohaemolyticus contains at least three serotypes, of which two have been defined in terms of agglutination reactions. Cross-agglutinations occurred between one strain of H. paraphrohaemolyticus and strains of the other V-dependent species, H. parainfluenzae and H. parahaemolyticus. Of the X-dependent species, H. haemoglobinophilus seems to be homogeneous, and the species H. aphrophilus is not. A non-specific antibody against horse blood in the medium occurred erratically and was present in only two antisera, those raised to H. aphrophilus strain Khairat and H. haemoglobinophilus strain no. NCTC8540.

Agglutination

Immunohistological analysis of intravascular agglutination in a case of ABO incompatible blood transfusion by means of mixed cell agglutination reaction.

Applicability of the mixed cell agglutination reaction (MCAR of Davidsohn) in histological examination was tested in a case of ABO incompatible blood transfusion, who had died of severe head injury two days after an accident. Massively disseminated intravascular aggregation of erythrocytes was identified as immune agglutination by the modified method of MCAR (ISHIYAMA & OKADA) with precise knowledge of topographic isoantigen localization in tissues. The immune specific agglutination mimics intravascular changes such as hyperemia, stagnation and hemorrhage derived from various circulatory disturbances obtained in routine autopsy materials.

ABO Blood-Group System

Cathepsin D agglutinators and neutral protease agglutinators in rheumatoid arthritis. II. Production of CDA and NPA by rheumatoid synovium and phagocytosis of CDA by synovial phagocytic cells.

Tissue culture methods demonstrated the production of agglutinators against the cathepsin D site in IgG (CDA) and a neutral protease site in IgG (NPA) by rheumatoid synovial tissue. Seven of 11 specimens from seropositive and 2 of 7 specimens from seronegative RA patients were positive for CDA, whereas 2 of 11 specimens from seropositive and 1 of 7 specimens from seronegative patients were positive for NPA. None of the 6 control specimens was positive for both types of agglutinators. Chromatography of two synovial tissue incubates showed that the CDA were of the IgG type. By the immunofluorescence technique, plasma cells containing CDA were demonstrated in rheumatoid synovial tissue and draining lymph nodes of rheumatoid joints. The staining for CDA in the synovium was different from the staining for pepsin agglutinators in adjacent sections. Phagolysosomes containing CDA were found in synovial exudate cells from rheumatoid patients as well as in phagocytosing lining cells and macrophages of the sublining layer of rheumatoid synovial tissue. These findings suggest that antibodies directed at hidden antigenic sites in IgG revealed by endogenous proteolysis take part in the immune reaction and in the inflammation of rheumatoid joints.

Agglutinins

Latex agglutination and adenoviruses. I. Detection of polyclonal antibodies by latex agglutination test.

A latex agglutination (LA) test was developed for the detection and measurement of polyclonal anti-adenovirus antibodies. Latex particles, coated with purified hexon, penton or fibre antigens displayed specific agglutination with 21 rabbit immune sera directed against the different antigens and the complete virus of 10 human and 2 animal adenovirus types. The sensitivity of LA was compared to that of passive haemagglutination, complement fixation (CF) and immunodiffusion reactions. In experiments with immune sera as well as with human serum samples the sensitivity of LA test proved to be in the same range as that of CF test. Because of its easy and rapid performance (slide agglutination within 4 min) LA method may substitute for CF reaction in serological adenovirus diagnosis.

Adenoviridae

Comparison of the agglutination and the co-agglutination techniques in T-tying of Streptococcus pyogenes.

A comparison of two different methods in T-typing 200 Norwegian group A strains is reported. The traditional agglutination technique was compared with the co-agglutination method of Christensen and co-workers. Fifty-five strains could not be typed by either method. The typing results for 128 of the strains were in agreement. The most common T types were the folloiwng: 1, 4, 8, 12, 13, 28, and strains belonging to the T pattern 5/11/12/27/44.

Agglutination Tests

Evaluation of five commercial tests: complement fixation, microparticle agglutination, indirect immunofluorescence, enzyme-linked immunosorbent assay and latex agglutination, in comparison to immunoblotting for Mycoplasma pneumoniae serology.

A panel of 68 serum specimens from 41 subjects exhibiting various immunological patterns to Mycoplasma pneumoniae as determined by detection of a 180 kDa protein in immunoblotting was used to compare five commercially available tests based on different methods: complement fixation test (CFT), microparticle agglutination (MAG), indirect immunofluorescence assay (IFA), enzyme-linked immunosorbent assay (Elisa), and latex agglutination (LA). The tests were performed according to the manufacturers' instructions. For the determination of immunity to M pneumoniae, the five tests were in good accordance with immunoblotting: sensitivity was 100% for all the five assays, specificity ranged from 95.6% (MAG) to 82.6% (Elisa) and overall agreement ranged from 98.2% (MAG) to 92.8% (Elisa). The comparisons of antibody rates obtained by the four quantitative tests (CFT, MAG, IFA, Elisa) showed correlation coefficients ranging from 0.87 (CFT-IFA) to 0.67 (CFT-Elisa). Six significant antibody rises demonstrated by immunoblotting patterns were detected by all the tests but Elisa in one case. As a whole, the commercial assays gave satisfactory results for routine determination of immune status to M pneumoniae: CFT was the cheapest test and MAG and LA were the easiest to perform.

Adolescent

[Highly specific agglutinating O- and OK-immunoglobulins for identification of Escherichia in the agglutination test on glass].

Escherichia agglutinating O- and OK-immunoglobulins of class G were obtained by the method of ion exchange chromatography; in studying with live and heated O- and K- cultures of the test strains from the International escherichia collection these immunoglobulins proved to be diagnostically highly specific and useful for the identification of escherichia strains in the express agglutination reaction on glass.

Agglutination Tests

Mating reaction in Saccharomyces cerevisiae. X. Agglutinability-inactivating factor: a factor which destroys sexual agglutinability of a mating-type cells.

From cells of Saccharomyces cerevisiae a factor has been extracted that destroys the agglutinability of a mating-type cells specifically. It was found in the cell extracts of diploid and tetraploid strains as well as haploid strains of a and alpha mating types. It is heat-labile and the molecular weight is about 50,000. It is adsorbed by neither a cells nor alpha cells. Its biological activity is dependent on the incubation temperature and the pH, and is completely inhibited by phenylmethylsulfonyl fluoride, a potent inhibitor of the serine proteases. All the results described in this paper indicate that this factor is a proteolytic enzyme.

Adsorption

Rapid identification of group A, B, C and G beta-haemolytic Streptococci by a modification of the co-agglutination technique. Comparison of results obtained by co-agglutination, fluorescent antibody test, counterimmunoelectrophoresis, and precipitin technique.

A rapid modification of the co-agglutination (COA) technique for grouping A, B, C and G beta-haemolytic streptococci was developed. The results are obtained within three hours after inoculation from primary plates of 0.5 ml broth. This method was compared to the fluorescent antibody test (FA) and counterimmunoelectrophoresis (CIE), two other rapid methods available for serological grouping of streptococci. Of 71 recently isolated streptococcal strains from clinical sources, 70 were correctly grouped in COA, 63 in FA and 68 in CIE. With commercial reagents COA compared favourably in accuracy to the other methods and can be recommended for routine serological grouping of beta-haemolytic streptococci in the clinical laboratory.

Agglutination Tests