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The agglutination of Proteus vulgaris by cystic fibrosis serum: a re-examination.

The agglutination of the bacterium Proteus vulgaris by serum from cystic fibrosis patients and obligate heterozygotes was shown to be insufficiently specific or reproducible to be of diagnostic value by itself. Approximately 20% of health controls gave a substantial agglutination reaction, whereas the carrier frequency for cystic fibrosis is around 5%. The agglutination did not predominantly involve the bacterial flagella, but appeared to depend on components of the cell surface. The main serum proteins that bind to P. vulgaris cells were shown to be albumin, immunoglobulin G, and complement component C3. In addition, an unidentified protein(s) of low molecular weight was found to bind to the cells. However, no systematic differences were found in the proteins that bind to P. vulgaris cells between cystic fibrosis and normal sera.

Agglutination↗

Simplified assay for concanavalin A-dependent bacterial agglutination by using cell surface mutants.

Two cell surface mutants of Escherichia coli isolated through supersensitivity to sodium dodecyl sulfate were found to be agglutinated by concanavalin A in the intact rod form. By using these strains, the assay method for the concanavalin A-dependent agglutination was greatly simplified since neither spheroplast formation nor protease treatment is needed. This rod cell agglutination was characterized and applied for a semiquantitative determination of the amount of bound concanavalin A.

Agglutination↗

TAXONOMY OF CLOSTRIDIUM BIFERMENTANS AND CLOSTRIDIUM SORDELLII. 3. AGGLUTINABILITY OF HEAT-RESISTANT SUBSTRAINS OFCLOSTRIDIUM SORDELLII.

Huang, C. T. (Kanazawa University, Kanazawa, Japan), Kenzo Tamai, and Shoki Nishida. Taxonomy of Clostridium bifermentans and Clostridium sordellii, III. Agglutinability of heat-resistant substrains of Clostridium sordellii. J. Bacteriol. 90:391-394. 1965.-By cross-agglutination tests with Clostridium bifermentans and C. sordellii antisera, the heat-resistant substrains from each pathogenic strain of C. sordellii were found to possess antigens of C. bifermentans in addition to displaying changes in biological activities. The agglutinability and biological activities varied with the sporulating potency acquired. Agglutinin-absorption tests further proved that heat-resistant substrains of C. sordellii were immunologically identical to C. bifermentans.

Agglutination↗

Mixed antiglobulin reaction and tray agglutination test for detection of sperm antibodies.

OBJECTIVE: To compare levels of sperm-agglutinating antibodies in male and female sera, cervical ovulatory mucus, and seminal plasma by two different tests. PATIENTS AND METHODS: 408 women and 404 men with long-lasting unexplained infertility and recurrent IVF failure were immunologically examined. Tray agglutination and mixed antiglobulin reaction tests were used for the comparison of sperm-agglutinating antibodies in sera, in ovulatory cervical mucus, and in seminal plasma. RESULTS: High secretory IgA positivity predominated in seminal plasma in 16 (3.9%) of infertile men and in cervical ovulatory mucus in 114 (27.9%) of infertile women. Positivity in two sperm antibodies together, in IgG and IgA, and IgM and IgE was also observed. Immunological anti-sperm antibodies in sera indicate high levels of sperm antibodies in IgG and IgM. CONCLUSION: Sperm immunoglobulin specification is important for therapeutic influence upon immunocompetent cells.

Agglutination Tests↗

A novel magnetic particle agglutination in microtiter plates for rapid detection of human T-lymphotropic virus type I antibody.

We have developed a novel magnetic particle agglutination (MPA) method for rapid detection of antibody to human T-lymphotropic virus type I (HTLV-I). This method is suitable for screening determination. The assay uses purified viral antigen coated on magnetic particles, which are novel artificial particles made of gelatin, arabic gum, and ferrite. MPA uses a special magnet and inclined plate holder. In MPA, the specimen is incubated with magnetic particles in the well; magnetic particles are then magnetically attracted to the bottom of the well. Then the plate is inclined and read by the naked eye. In a positive specimen, agglutination particles stay at the bottom of the well. On the other hand, non-agglutination particles with negative sample run down the side of the well. The total assay time of MPA was estimated at 8 min/microtiter-plate. The results obtained by correlating MPA for the detection of HTLV-I antibody with other method emphasized the precision of MPA.

Adult↗

The effects of neuraminidase on concanavalin A agglutination of erythrocytes: evidence for adsorption of neuraminidase to erythrocyte membrane.

Neuraminidase-treated human erythrocytes, but not untreated erythrocytes, were agglutinated by concanavalin A. The degree of concanavalin A agglutinability was not directly related to sialic acid removal by neuraminidase. While maximal sialic acid release was obtained with 5 units neuraminidase/2 x10(9) erythrocytes, maximal concanavalin A agglutination was only obtained after exposure to 20 units neuramindase. Binding of 3H-concanavalin A by erythrocytes was 10-fold higher with rabbit compared to human red cells. Neuraminidase treatment of human erythrocytes caused a relative increase in 3H-concanavalin binding, but the absolute amount was still 10-fold less than that bound to rabbit erythrocytes. Specific adherence of neuraminidase to Con A-Agarose could not be demonstrated. There was no evidence for contamination of the neuraminidase preparation with proteases using a sensitive assay. These studies suggest that neuraminidase absorbs to erythrocytes by a mechanism other than removal of sialic acid.

Adsorption↗

The dissociation of the surface architecture described by enhanced lectin agglutinability and the transformed phenotype expressed as anchorage independence.

Using a series of cold-sensitive variants of chemically transformed BHK-21 cells, revertants to the normal phenotype derived from a dimethyl-nitrosamine transformed clone of BHK-21 as well as revertants to the normal phenotype derived from polyoma transformed BHK-21 cells we have demonstrated that the surface phenotype described by enhanced agglutinability with Con A and WGA can be dissociated from the transformed phenotype described by anchorage independence (growth in semisolid medium). Specifically we have demonstrated that the surface characteristic of enhanced agglutinability may be found in a variety of cell lines which fail to display to grow in agar. Our work clearly shows that the two phenotypes described are not concomitantly controlled and tends to suggest that the phenotype of enhanced lectin agglutinability may be dissociated from the transformed phenotype.

Agar↗

Carcinogenesis or tumourigenicity testing of animal cell lines for vaccine preparation by colony formation on soft agar and by agglutination under plant lectins.

The carcinogenic or tumourigenic testing of seven animal kidney cell lines (F-81, CRFK, MDCK, Vero, Vero-2 cell line, MA-104 and BHK-21) established in China, were carried out in more than 700 nude mice for colony formation in soft agar and for agglutination under different density of plant lectins. Tests showed that there were correlation between cell line chromosome number variations and anchorage independence in soft agar, agglutinability under lectins and tumour-forming ability in nude mice. Since testing in vitro was more economical, simpler and faster and thus thought to be more reliable, we recommend measuring agglutinability, followed by anchorage independence or analysis of karyotype as the initial means for monitoring tumourigenicity of animal cell lines in nude mice.

Agar↗

Increased agglutinability of bladder epithelial cells by concanavalin A in rats fed several biphenyl derivatives.

Biphenyl and its derivatives, 2-aminobiphenyl, 2-nitrobiphenyl, 4-aminobiphenyl, 4-nitrobiphenyl, p-phenylphenol, o-phenylphenol (OPP), and o-phenylphenol sodium salt tetrahydrate (OPP-Na) were examined for their bladder carcinogenicity in rats by a short-term assay for agglutinability of bladder epithelial cells with concanavalin A. Increased agglutinability was observed after 1-week treatment with 2.0% and 1.0% OPP, 2.0% and 1.0% OPP-Na, 0.5 and 0.1% 4-aminobiphenyl, and 0.5% 4-nitrobiphenyl in the diet, suggesting carcinogenicity of these compounds in the bladder. Such an increase in agglutinability was not observed in rats fed diets containing biphenyl, 2-aminobiphenyl, 2-nitrobiphenyl or p-phenylphenol at 2.0%. With OPP-Na, an in vivo carcinogenesis experiment was performed. Bladder carcinomas developed in 14 of 36 male Fisher rats fed a 2% OPP-Na diet for 50 weeks.

Agglutination Tests↗

P-fimbriae of pyelonephritogenic Escherichia coli: detection in clinical material by a rapid receptor-specific agglutination test.

A slide agglutination test to detect P-fimbriated Escherichia coli was developed by attaching glycosides containing the relevant receptor structure (alpha-D-Galp-(1-4)-beta-D-Galp) to particles. A suspension of these particles agglutinates within ten seconds when mixed with P-fimbriated bacteria. The test is named the P-specific particle agglutination test (PPA test). The PPA test is more sensitive than the haemagglutination assays previously used. The exclusive specificity of the PPA test also allows the detection of P-fimbriae on strains which additionally possess other fimbriae (e.g. X-fimbriae). Using this test, the frequency of P-fimbriated E. coli in children with acute non-obstructive pyelonephritis was shown to exceed 95%. In lower urinary tract infections the frequency of P-fimbriated E. coli was approximately 20%. Moreover, children with acute pyelonephritis were also found to be heavily colonized in both the periurethral area and the intestine with the identical P-fimbriated E. coli strain.

Adult↗

Microparticle agglutination versus antibody-capture enzyme immunoassay for diagnosis of community-acquired Mycoplasma pneumoniae pneumonia.

Community-acquired Mycoplasma pneumoniae pneumonia is a common disease which is usually diagnosed by serological methods. The objective of the present study was to understand the diagnostic significance and test characteristics of two different serological tests used to identify current Mycoplasma pneumoniae infection. Three hundred sixty-six patients who suffered from community-acquired pneumonia served as the study population. Six hundred ninety-four (328 paired and 38 unpaired) sera were examined for the presence of antibodies to Mycoplasma pneumoniae with commercial kits based on two serological methods, microparticle agglutination and antibody-capture EIA. Agreement between the two kits was 85.2% when individual sera were compared (kappa = 0.62) and 88.5% when patients were compared (Kappa = 0.69). The positive predictive value and the specificity for the identification of current Mycoplasma pneumoniae infection using a single acute-phase serum were 49.3% and 86.9%, respectively, for the microparticle agglutination method, compared to 91.3% and 97.7% for the antibody-capture EIA method (p < 0.001). The negative predictive value and the sensitivity were 86.3% and 48.1% for the microparticle agglutination, not significantly different from the corresponding values of 86.5% and 61.2% for the antibody-capture EIA. It is concluded that the overall agreement between the two methods tested is good, but not perfect. The methods complement each other in the identification of Mycoplasma pneumoniae as the causative agent in patients with community-acquired pneumonia.

Adolescent↗

Agglutinins from aquatic insects--tumor cell agglutination activity.

Agglutinins were identified in whole body extracts of aquatic insects by means of murine tumor cell agglutination, using sarcoma 180 ascites, Ehrlich, and MM-46 cells. Screening revealed agglutinins in 5 of 10 of the larvae tested, and in 2 of 6 of the water-dwelling adult insects; Gerris paludum insularis and Gyrinus japonicus. Only the agglutinin from adult G. paludum also agglutinated human erythrocytes. An ascites tumor was converted into a solid form in vivo after administration of G. paludum agglutinin. The observation that these aquatic insect agglutinins preferentially agglutinate tumor cells has considerable implications in terms of anti-tumor effects such as inhibition of cell proliferation and metastasis.

Agglutination↗

Synthetic identification as a hexapeptide of alpha substance-IB inducing sexual agglutination in Saccharomyces cerevisiae.

For the identification of a peptidyl principle inducing sexual agglutination in the yeast, 2 supposed hexapeptides (1a, b) were synthesized by the conventional method. The 1a (H-Arg-Gly-Pro-Phe-Pro-Ile-OH) revealed complete identity with the natural peptide in TLC, MS and biological property on agglutination. The 1b showed the sexual agglutinability in the same degree as 1a, though distinct differences were observed in the chemical data. Both 1a and 1b had a strong bitter taste.

Agglutination↗

Comparison of enzyme-linked immunosorbent assay with coagglutination and latex agglutination for rapid diagnosis of pneumococcal pneumonia by detecting antigen in sputa.

A sandwich enzyme-linked immunosorbent assay detecting the species-specific pneumococcal C polysaccharide was compared to latex agglutination and a coagglutination test which detected capsular pneumococcal antigens in sputum specimens with regard to specificity and sensitivity. Specimens from 52 patients with clinical and radiological evidence for pneumonia were tested. Twenty-one patients with Streptococcus pneumoniae isolated in sputum and 31 patients with a non-pneumococcal etiology were included. The predictive values for a positive test by enzyme-linked immunosorbent assay was 0.91 and for a negative test 0.97, by latex agglutination 0.90 and 0.91, and by coagglutination 0.84 and 0.85 respectively; these values did not show a statistically significant difference. Whereas agglutination tests are technically more simple and can be performed more rapidly, the enzyme-linked immunosorbent assay has the advantage of detecting pneumococcal C polysaccharide, an antigen common to all pneumococci. Thus it provides an interesting alternative to tests based on serum containing antibodies to all 83 different capsular polysaccharides.

Adolescent↗

Direct agglutination test for serologic diagnosis of Neospora caninum infection.

A direct agglutination test was evaluated for the detection and quantitation of IgG antibodies to Neospora caninum in both experimental and natural infections in various animal species. As compared with results obtained by the indirect fluorescent antibody test, the direct agglutination test appeared reliable for the serologic diagnosis of neosporosis in a variety of animal species. The direct agglutination test should provide easily available and inexpensive tools for serologic testing for antibodies to N. caninum in many host species.

Abortion, Veterinary↗

An analysis of concanavalin A-mediated agglutination in two Chinese hamster ovary subclones whose surface phenotypes respond to maintenance in medium supplemented with dibutyryl cyclic AMP. V. Biochemical composition of the plasma membrane.

We have used two Chinese hamster ovary subclones whose surface phenotype has been extensively investigated with regard concanavalin A-mediated cell-cell agglutination and concanavalin A-induced receptor site clustering to investigate what changes in membrane composition, if any, can be correlated with the concanavalin A-detected changes in surface phenotype. These cell clones are uniquely disposed for this purpose since maintenance of the cells under different growth conditions produces changes in agglutinability and receptor site mobility in one cell clone (H-7W) but not the other (K-1). After extensive characterization of the surface membranes of these two subclones we have been unable to identify any change in the membrane peptides, glycopeptide, cholesterol, or fatty acid composition which can be directly correlated with the concanavalin A-detected surface phenotypes. It is of particular interest to note that we have been unable to correlate the presence or absence of the large external transformation-sensitive glycoprotein with the relative mobility of the lectin receptors or with the degree of concanavalin A-mediated cell agglutination. Furthermore we have been unable, in this system, to corroborate earlier data suggesting a role for cholesterol in determining the relative mobility of the lectin receptors. Thus using a cell system consisting of genetically matched cell clones, we have been unable to identify any changes in the biochemical composition of the plasma membrane which might be associated with the surface phenotypes detected by concanavalin A.

Agglutination↗

Agglutination microassay of hapten- or protein-modified liposomes using a multiple cell culture harvester.

A simple agglutination microassay is described for studying the presence of antigenic determinants on the surface of liposomal membranes. Antigen-sensitized radioactive liposomes, reacted with specific immune IgG in the wells of a microtest plate, were passed through a glass fiber filter in a multiple cell culture harvester. The amount of agglutinate trapped on the filter as determined by the radioactivity, was proportional to the amount of antigen in the liposomes and to the amount of specific immune IgG in the reaction mixture. The specificity of the agglutination was determined by inhibition with hapten and carrier antigens. Liposomes actively sensitized with hapten-conjugated lipid, and passively sensitized with protein, were used in the assay. A new method of preparing protein-bound liposomes is also described.

Agglutination Tests↗