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PubMed1955

Specific agglutination of Treponema pallidum by sera from rabbits and human beings with treponemal infections.

A method has been described for the preparation of Treponema pallidum suspensions that are suitable for specific agglutination studies and can be stored at 4 degrees C. for months without loss of agglutinability. Such suspensions have been shown to react with two distinct antibodies in the serum of syphilitic animals and man: Wassermann antibody and a specific treponeme agglutinin. It has been demonstrated that the agglutination of treponemes by specific treponeme agglutinin is enhanced by heat treatment or aging of the suspension, and inhibited by a divalent cation, probably Ca(++), normally present in serum. This inhibition has been overcome by the use of a chelating agent, ethylene-diamine tetracetate. These findings have been utilized to devise a simple agglutination test for the diagnosis of treponeme infections that is very sensitive and highly specific. This test has been carried out with 430 human sera, and a comparison has been made of the results of the agglutination, treponemal immobilization, and standard serological tests on these sera. The agglutination test appears to have a specificity comparable to the treponemal immobilization test and considerably greater than the standard serological tests.

Agglutination↗
PubMed1992

An analysis of lectin agglutination as a means of sub-dividing gonococcal serovars.

Sixteen lectins were examined for their ability to agglutinate 298 strains of Neisseria gonorrhoeae. Seven lectins failed to agglutinate any of the strains; the remaining nine lectins gave 22 different agglutination patterns. The 298 strains were divided into 14 serovars with a single panel of monoclonal antibody typing reagents; lectin agglutination subdivided these into 57 serovar/lectin patterns. A combination of two monoclonal antibody serotyping panels divided the strains into 32 serovar combinations; lectin agglutination further subdivided these into 79 serovar/lectin patterns. There was no correlation between lectin pattern and serovar. Lectin agglutination is a simple supplementary typing method and could be particularly useful in micro-epidemiological studies.

Agglutination↗
PubMed1994

Agglutination of "Streptococcus milleri" by lectins.

The agglutination of 218 clinical isolates and three ATCC type strains of "Streptococcus milleri" was tested with 25 different lectins from plants and fungi. An agglutination reaction with one or more lectins was observed with 42 isolates when the cells were untreated. After trypsinisation of the bacteria, 109 strains yielded a positive reaction and after boiling the bacterial cells at pH2, 218 isolates were agglutinated. As an overall result of our experiments with untreated, trypsinised and boiled cells, 17, 37 and 45 different agglutination patterns, respectively, were obtained. The lectins from Datura stramonium, Robinia pseudoacacia and Dolichos biflorus agglutinated isolates belonging only to Lancefield group C, being non-reactive with other isolates. These lectins were also found to be specific for "large colony type" streptococci of group C. The use of lectin agglutination in epidemiological and ecological studies of "S. milleri" is discussed.

Agglutination↗
PubMed1962

Fluorescent antibody staining and agglutination reactions in Bordetella pertussis cultures.

Eldering, Grace (Michigan Department of Health, Grand Rapids), Warren C. Eveland, and Pearl L. Kendrick. Fluorescent antibody staining and agglutination reactions in Bordetella pertussis cultures. J. Bacteriol. 83:745-749. 1962-Bordetella pertussis antisera produced with smooth cultures gave positive results in agglutination and fluorescent antibody (FA) tests with smooth B. pertussis cultures. Similar results were obtained with antisera produced with the related species B. parapertussis and B. bronchiseptica when each was tested with the respective homologous antigen. Cross reactions occurred with some of the antisera and heterologous antigens but a positive agglutination test was not always correlated with a positive FA reaction, and the reverse was also observed. Adsorbed B. pertussis antisera specific for factors 2, 3, 4, and 5 agglutinated appropriate B. pertussis cultures, but gave negative FA reactions. Factor 1 antiserum prepared by adsorption with heated (100 C) antigen gave positive results in both FA and agglutination tests. Rough B. pertussis cultures were inagglutinable, but were FA positive when tested with unadsorbed antiserum produced with either smooth or rough cultures. Rough cultures were FA negative with factor 1 serum. The results suggest that for B. pertussis different serum components may be responsible for agglutination and FA staining. The implications with respect to the protection-inducing antigen are discussed.

Agglutination↗
PubMed1975

Difficulty of obtaining reproducibility in the Franklin and Dukes test for the detection of sperm-agglutinating antibodies in human sera.

The Franklin and Dukes (FD) test is used as a screening test for the determination of sperm-agglutinating antibodies and as a proof of therapeutic effects in cases of infertility, but various parameters of the FD test remain unclear and the application of different techniques has revealed highly different results. To test the statistical evidence for the FD technique, conditions regarded as optimal prerequisites were applied: a sperm concentration of 20 times 10(6)/ml was produced with Baker's buffer and serum dilutions were made with Baker's buffer, starting at 1:4. Only the standard deviation of parallel tests with one serum sample and one semen sample on the same day was found to be within an acceptable range. Using ejaculates of the same donor and the same serum sample on different days gave results that were not reproducible. The FD test should not, therefore, be used for quantification of sperm-agglutination antibodies except for a comparison in one test with one semen sample on the same day. Although the FD test only allowed a qualitative evaluation of sperm-agglutinating antibodies the % sperm agglutination was more informative than agglutination titres. For quantification of sperm-agglutinating antibodies, the FD test should be replaced by other techniques.

Agglutination Tests↗
PubMed1979

Sperm-agglutinating antibodies in infertile couples.

In a selected group of 150 infertile couples, sperm-agglutinating antibodies were examined by a microagglutination technique. Sperm-agglutinating activity was observed in sera from 12 men (8%) and 16 women (11%). Antibodies in seminal plasma were demonstrated in 8 men with correspondingly high serum titers. The type of agglutination in these cases was head to tail. Head-to-head agglutination was found in only 3 men, who all had agglutinating activity in serum only. An abnormal postcoital test was found in 57% of the couples, and 13% of the men and 14% of the women in these couples had agglutinating antibodies. A negative postcoital test was related more often to high than to low titer in female sera.

Agglutination Tests↗
PubMedDate not supplied

The effect of oil-emulsion vaccines on the occurrence of nonspecific plate agglutination reactions for Mycoplasma gallisepticum and M. synoviae.

Six groups of ten 18-week-old mycoplasma-free white leghorn pullets were vaccinated with one of the following: Mycoplasma gallisepticum (MG) bacterin. Haemophilus gallinarum bacterin, Pasteurella multocida bacterin, combined infectious bursal disease (IBD)-Newcastle (NDV) chicken-embryo-origin (CEO) vaccine. IBD-NDV tissue-culture-origin (TC) vaccine, or saline emulsified in oil; one group received no vaccine. Plate agglutination tests for M. synoviae (MS) and MG were done for 10 weeks after vaccination using three different test antigens. Pullets vaccinated with H. gallinarum bacterin and IBD-NDV TC vaccine showed the greatest incidence of nonspecific plate agglutination reactions. The incidence of positive plate agglutination reactions varied with test antigens. Five groups of fifty 18-week-old mycoplasma-free heavy-breed pullets were vaccinated with one of the following: saline emulsified in oil, chicken embryo fibroblasts emulsified in oil, allantoic fluid emulsified in oil, chicken embryos emulsified in oil, or MS-contaminated chicken embryos emulsified in oil. Plate agglutination tests for MS and MG were done for 8 weeks after vaccination. Chickens vaccinated with chicken embryo fibroblasts emulsified in oil had the greatest incidence of nonspecific plate agglutination reactions. Pullets vaccinated with MS-contaminated chicken embryo vaccine had only a small increase in MS-positive plate agglutination reactions compared with pullets vaccinated with uncontaminated chicken embryo vaccine.

Adjuvants, Immunologic↗
PubMed1985

Mannose inhibition of Escherichia coli adherence to urinary bladder epithelium: comparison with yeast agglutination.

The adherence of piliated strains of Escherichia coli (E. coli) to mammalian epithelial cells has been reported by several investigators to be specifically inhibited by D(+)-mannose or its derivatives. Much of this work utilized mannose type compounds to inhibit agglutination of mannan containing yeast cells by E. coli to demonstrate mannose sensitivity. This report investigates the ability of the neotype strain of E. coli (which is sensitive to mannose inhibition of yeast cell agglutination) to bind and metabolize radiolabeled D(+)-mannose. In addition the relative efficacy of D(+)-mannose and heparin to inhibit the adherence of E. coli to rabbit bladder mucosa was compared. Results showed that although D(+)-mannose did block E. coli--yeast cell agglutination in a reversible manner, radiolabeled D(+)-mannose binding by E. coli could not be displaced by 1,000 fold excess unlabeled D(+)-mannose. This suggests uptake of the sugar as opposed to a surface binding phenomenon which was confirmed by the demonstration of significant metabolism of mannose by E. coli. The same concentration of D(+)-mannose which prevented E. coli--yeast cell agglutination was not particularly effective in preventing E. coli adherence to the acid denuded rabbit bladder. Heparin treatment of the acid denuded bladder was very effective in preventing E. coli adherence but was ineffective in preventing E. coli--yeast cell agglutination. This indicates that E. coli--yeast cell agglutination should not be correlated with E. coli adherence to mammalian epithelial tissue.

Agglutination↗
PubMed1989

Genetic analysis of inducible sexual agglutination ability in the yeast Saccharomyces cerevisiae.

Genetic regulation of the inducibility of sexual agglutination ability in the yeast Saccharomyces cerevisiae was studied. Detailed analysis of the degree of sexual agglutination was carried out; it showed that a greater number of genes are involved in the regulation of inducible sexual agglutination in strain H1-0 than previously assumed. Although dominancy of inducible phenotype over constitutive was confirmed, the effectiveness of one gene changing the constitutive phenotype to the inducible seemed to be somewhat low. Quantity per cell of agglutination substances responsible for sexual agglutination increased as the agglutination ability became greater.

Agglutination↗
PubMed1976

Mating reaction in Saccharomyces cerevisiae. IX. Regulation of sexual cell agglutinability of a type cells by a sex factor produced by alpha type cells.

A diffusible sex-specific substance called alpha substance-I (alphaS-I) was isolated from culture filtrate of alpha type strains of the yeast Saccharomyces cerevisiae. The isolated alphaS-I, an oligopeptide, induced sexual cell agglutinability in inducible alpha type strains and enhanced the agglutinability in constitutive a type strains. The induction of sexual agglutinability was detected in 30 min and reached maximum in 90 min, when 0.2 mug/ml of alphaS-I was added to inducible a type cells. The a type-specific factor responsible for sexual cell agglutination, called a type agglutination factor (aAF), was shown to be produced during the induction or the enhancement of agglutinability of a type cells by alphaS-I. The aAF produced in response to alphaS-I was not different in the susceptibility to proteolytic enzymes and disulfide-cleaving agents from those produced constitutively in the absence of alphaS-I.

Agglutination↗
PubMed1975

Effects of local anesthetics on membrane properties. II. Enhancement of the susceptibility of mammalian cells to agglutination by plant lectins.

Treatment of untransformed mouse and hamster cells with the tertiary amine local anesthetics dibucaine, tetracaine and procaine increases their susceptibility to agglutination by low doses of the plant lectin concanavalin A. Agglutination of anesthetic-treated untransformed cells by low doses of concanavalin A is accompanied by redistribution of concanavalin A receptors on the cell surface to form patches, similar to that occurring in spontaneous agglutination of virus-transformed cells by concanavalin A. Immunofluorescence and freeze-fracture electronmicroscopic observations indicate that local anesthetics per se do not induce this redistribution of concanavalin A receptors but modify the plasma membrane so that receptor redistribution is facilitated on binding of concanavalin A to the cell surface. Fluorescence polarization measurements on the rotational freedom of the membrane-associated probe, diphenylhexatriene, indicate that local anesthetics produce a small increase in the fluidity of membrane lipids. Spontaneous agglutination of transformed cells by low doses of concanavalin A is inhibited by colchicine and vinblastine but these alkaloids have no effect on concanavalin A agglutination of anesthetic-treated cells. Evidence is presented which suggests that local anesthetics may impair membrane peripheral proteins sensitive to colchicine (microtubules) and cytochalasin-B (microfilaments). Combined treatment of untransformed 3T3 cells with colchicine and cytochalasin B mimics the effect of local anesthetics in enhancing susceptibility to agglutination by low doses of concanavalin A. A hypothesis is presented on the respective roles of colchicine-sensitive and cytochalasin B-sensitive peripheral membrane proteins in controlling the topographical distribution of lectin receptors on the cell surface.

Agglutination↗
PubMed1977

The effect of temperature on concanavalin A-mediated agglutination of cells with rigid receptors.

The agglutination of a yeast, Candida albicans, by concanavalin A has been described. The agglutination was cell-number dependent. Prolonged incubation (60 min) was needed to reach maximum agglutination at 37 degrees C. The rate but not the extent of agglutination was temperature dependent. The dimeric forms of concanavalin A, obtained either at low pH or after succinylation, agglutinated the yeast cells as well as the tetramer. Temperature changes affected the agglutination of yeast cells by dimers and by tetramers to the same extent.

Agglutination Tests↗
PubMed1979

Membrane lipid fluidity as rate limiting in the concanavalin A-mediated agglutination of pyBHK cells.

The initial rate of concanavalin A-mediated agglutination of polyoma transformed Baby Hamster Kidney (pyBHK) cells follows Arrhenius kinetics. There is a smooth decrease in the agglutination rate from 37 degrees C to 22 degrees C with an activation energy of 11.8 +/- 0.2 kcal/mol in this region. There is a sharp decrease in agglutination rate below 22 degrees C. The addition of 0.1 mM 1,3-di-tert-2-hydroxyl-5-methylbenzene, a lipid perturber, increases the agglutination rate by a factor of two and increases the membrane lipid fluidity as determined by the spin label method. The rotational correlation time of the spin label 2N14 (2,2-dimethyl-5-dodecyl-5-methyloxazolidine-N-oxide) was measured. The sum of the enthalpy of activation of rotational diffusion and the enthalpy of activation of translational diffusion is very nearly equal to the enthalpy of activation of agglutination. This is consistent with the rate limiting step of agglutination being receptor diffusion, which is probably limited in pyBHK cells by membrane lipid fluidity.

Agglutination↗
PubMed1991

Fractionation of jelly substance of the sea urchin egg and biological activities to induce acrosome reaction and agglutination of spermatozoa.

Jelly coat substance surrounding the egg of the sea urchin, Hemicentrotus pulcherrimus, was fractionated by gel filtration and three fractions designated A, B, and C were obtained which mainly consisted of fucose sulfate, sialic acid, and protein, respectively. The biological activities of the fractions were examined for induction of acrosome reaction (AR) and agglutination of spermatozoa. Only fraction A, a fucose-rich glycoprotein fraction, had activities for both AR and agglutination. Fraction A was found to lose activity for AR but to retain activity for agglutination after pronase digestion. Pronase-digested fraction A was further fractionated by the same gel filtration and three fractions designated P1, P2, and P3 were obtained, which contained mainly fucose sulfate, sialic acid, and proteinous material, respectively. These fractions had no activity for AR but activity for agglutination resided in fraction P1, a fucose sulfate fraction. Furthermore, beta-elimination of the jelly substance was carried out to separate protein and fucose sulfate polysaccharide and three fractions designated E1, E2, and E3 were obtained by gel filtration, of which the fucose-rich fraction (E1) exhibited activities for both AR and agglutination, and the sialoprotein fraction (E2) retained activity only for AR. However, the activity for AR of both fractions was destroyed by pronase digestion. These results suggest that activity to induce AR resides in the protein moiety of fucose-rich glycoprotein and activity for agglutination resides in the fucose sulfate polysaccharide moiety of the same glycoprotein of the jelly substance.

Acrosome↗
PubMed1990

Immunological agglutination kinetics of latex particles with physically adsorbed antigens.

The influence of the properties of antigens and particles on the immunological agglutination kinetics of the antigen-coated latex particles was studied. Horse cytochrome c, hen egg-white lysozyme (HEL), bovine serum albumin (BSA), and Aspergillus sp. glucose oxidase were physically adsorbed onto the surfactant free latices of styrene-methacrylic acid (MAA) copolymer (P (S/MAA)) and polystyrene (PS). The initial rates of the immunological agglutination of these protein-coated particles initiated by the addition of antibodies were quantified by the absorbance change at a wavelength of 680 nm. The initial agglutination rates of the particles covered with smaller antigens were lower. This effect of the molecular size of antigens was larger in P(S/MAA), because small antigens are probably buried in the hydrous polymethacrylic acid layer on the surface of particles. Thus, both the molecular size of antigens and the surface properties of particles affect the sensitivity of the immunological agglutination. On the other hand, the dependence of the initial rate of the immunological agglutination on the ionic strength and pH was similar irrespective of antigen-particle systems. The initial agglutination rates were largest at an ionic strength of approximately 0.05 at pH 7.0 and decreased with increasing pH. This dependence of the sensitivity on the pH and ionic strength is attributed to the electrostatic interactions of particle-particle and antibody-particle.

Adsorption↗
PubMed1992

Detection of fecal blood by colloidal gold agglutination using an anti-human hemoglobin monoclonal antibody.

Monoclonal antibodies to human hemoglobin were produced and a colloidal gold agglutination method has been developed for detection of fecal occult blood. Since hemoglobin is composed of the tetramer, alpha 2 beta 2, a single monoclonal antibody-labeled colloidal gold can agglutinate with hemoglobin. The lowest detectable hemoglobin concentration was 0.5 micrograms/ml. A total of 785 fecal samples were determined using colloidal gold agglutination and compared with latex agglutination. The colloidal gold agglutination detected blood in 75 samples, whereas latex agglutination detected blood in 76 samples, and among them 70 were positive in both methods. Overall agreement between the two methods was 98%.

Agglutination↗
PubMed1991

Agglutination of bacteria and erythrocytes by serum from six species of marine molluscs.

Lectins have been implicated as recognition molecules in the invertebrate immune response, yet their capacity to recognize (agglutinate and/or opsonize) potentially invasive microorganisms is largely unknown. In this study, sera from six species of marine molluscs (oyster, clam, octopus, squid, cuttlefish, and sea hare) were found to agglutinate 64 of 94 bacterial isolates (15 genera, 36 species) and seven types of vertebrate erythrocytes. Oyster, clam, and octopus sera agglutinated the greatest number of bacterial isolates and oyster serum exhibited the highest intensity and titer of agglutination. Agglutination was isolate dependent, implying high lectin specificity. Titers were highly variable for wild populations of oysters and sea hares and relatively constant for F4-generation cuttlefish reared in aquaculture systems. Simultaneous agglutination of specific bacterial isolates indicated conservation of lectin specificity between oyster and clam sera (bivalves) and between squid and cuttlefish sera (cephalopods).

Agglutination↗
PubMed1988

A rabies agglutination test (RAT) for rabies antibody detection.

An agglutination test has been developed for the detection of rabies antibodies after human vaccination. The rabies agglutination test (RAT) is based on the capability of specific antibody to agglutinate sensitized polystyrene (or latex) beads. In the RAT, latex beads were coated, in a first step, with inactivated and purified rabies virus (PV strain adapted and propagated on BHK-21 cells) and, in a second step, with bovine serum albumin. Negative control beads were coated with bovine serum albumin (BSA) only. To test for human antibody, several microliters of serum were mixed on a glass slide with an equal volume of virus-sensitized beads and the mixture was gently agitated. After a few minutes, agglutination was visible with sera which had been characterized as positive by the virus neutralization antibody (VNAb) technique. No agglutination was observed with negative sera tested with virus-coated beads or with positive sera tested with BSA-coated beads. Virus-sensitized beads were agglutinated when the virus neutralizing antibody titres were equal to or greater than 2.5 international units per ml (IU/ml) in human sera. The concordance between the RAT results and VNAb titres was about 97% when 2.5 IU/ml was taken as the cut off value for determining the positive sera with the VNAb technique. The possibility that clinicians might use the RAT as a simple means to determine sero-conversion at the end of the post-exposure treatment of patients is discussed.

Agglutination Tests↗