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PubMed1981

An Amphioxus lanceolatus agglutinating factor for human red cells.

Humoral proteins from Amphioxus lanceolatus showed a considerable agglutinating activity for human red cells. The haemagglutinating factor eluted in the macroglobulin peak on Sephadex G200 and resulted ConA-1 at affinity chromatography on Sepharose ConA. Immunoelectrophoresis, carried out using a specific antiserum produced in rabbit, revealed alpha mobility for the agglutinating factor, thus suggesting that it was an alpha macroglobulin.

Animals↗
PubMed1991

Detection of antiplatelet antibody: comparison of platelet immunofluorescence, agglutination, and immunoinjury tests using rabbit antiequine platelet serum.

Immunofluorescence, tube agglutination, and platelet factor-3 immunoinjury tests for detecting antiplatelet antibody were compared using a heterologous system of equine platelets and rabbit antiequine platelet serum. Platelet immunofluorescence tests were performed using paratormaldehyde-fixed platelets in suspension as well as in air-dried smears on glass slides (solid phase). Bright homogeneous, membranous, specific fluorescence was seen in both assays with anti-immunoglobulin G (IgG) and protein G fluorescein isothiocynate conjugates (FITC-conjugates). Protein A conjugate gave nonspecific fluorescence irrespective of normal or antiserum treatment. Anti-IgG and protein G conjugates in suspension immunofluorescence tests with the same antiserum yielded antibody titers of 1:1024 and 1:128, respectively. Similarly, respective titers of 1:512 and 1:64 were obtained with solid phase immunoassay. Platelet suspension assay was slightly better than the solid phase assay. These observations indicated that anti-IgG was more sensitive than protein G in detecting antiplatelet antibody by fluorescence microscopy, while protein A was ineffective because of its nonspecificity. Chloroquine treatment of platelets failed to reduce the nonspecific fluorescence. Platelet agglutination and platelet factor-3 tests were relatively less sensitive to detect equine antiplatelet antibody.

Journal Article↗
PubMed1981

Evaluation of a new latex agglutination inhibition card test for determining serum gentamicin levels.

A new latex agglutination inhibition card test was compared with a microbiological method and radioimmunoassay for determining serum gentamicin levels. When 60 sera, each containing a known concentration of gentamicin, were analyzed by the microbiological method and by radioimmunoassay, both methods were found to yield accurate and precise results. Analysis of the same 60 sera with the latex agglutination inhibition card test revealed significant variability over a wide range of serum gentamicin concentrations.

Bacillus↗
PubMed1988

Rapid latex agglutination test for detection of staphylococcal enterotoxins A to E that uses high-density latex particles.

A rapid reversed passive latex agglutination method that uses high-density latex particles for the detection of staphylococcal enterotoxins (SE) A to E was developed. It took 3 h for incubation, much less than the 16 h needed with a customary latex agglutination test for SE detection such as a commercial test kit (SET-RPLA; Denka Seiken Co. Ltd., Tokyo, Japan). The rapid test was shown to be highly specific and sensitive for SE detection (detection limit, about 0.5 ng of SE per ml), comparable to the SET-RPLA test. The rapid test was also efficient in SE detection in foods and culture supernatants of staphylococcal strains, similar to the SET-RPLA test. This showed that a rapid test with high-density latex particles is fully reliable for use.

Enterotoxins↗
PubMed1988

Fusarium Wilt Suppression and Agglutinability of Pseudomonas putida.

Mutants of Pseudomonas putida (Agg) that lack the ability to agglutinate with components present in washes of bean and cucumber roots showed limited potential to protect cucumber plants against Fusarium oxysporum f. sp. cucumerinum. However, a higher level of protection was observed against Fusarium wilt in cucumber plants coinoculated with the parental bacterium (Agg), which was agglutinable. The Agg mutants did not colonize the roots of cucumber plants as extensively as the Agg parental isolate did. In competition experiments involving bean roots inoculated with a mixture of Agg and Agg bacteria, the Agg strains colonized roots to a greater extent than the Agg cells did. These data suggest that the Agg phenotype provides additional interactions that aid in the beneficial character of P. putida.

Journal Article↗
PubMed1990

Screening of aquatic samples for Vibrio cholerae serotype O1 by a dot-blot method and a latex agglutination test.

A dot-blot, enzyme-linked immunosorbent method and a latex agglutination test were studied for their abilities to detect Vibrio cholerae serotype O1 in aquatic samples by testing artificially contaminated water as well as samples from natural potential sources. Water samples were preenriched with alkaline peptone and then enriched with Monsur peptone water. For the dot-blot test, enriched cultures of organisms in a small portion of the Monsur peptone water were transferred to a polyvinylidene difluoride membrane with a microfiltration apparatus. The enzyme-linked immunosorbent assay was performed by using biotin-labeled antibodies and avidin-biotin-peroxidase complex; brown dots developed in the wells that contained serotype O1 vibrios. Latex agglutination tests were performed by mixing 1 drop of the culture in Monsur with 1 drop of reagent coated with monoclonal antibody specific for antigen A. The sensitivities and specificities of the methods were compared with those of the colony-blot method, which identified individual colonies of V. cholerae O1 in mixed bacterial cultures on isolation media. Our results indicate that the dot-blot method is as sensitive as the colony-blot method and is useful for screening for V. cholerae serotype O1 even in specimens that are heavily contaminated with non-O1 vibrios.

Humans↗
PubMed1995

Biochemical Characterization and Agglutinating Properties of Xenorhabdus nematophilus F1 Fimbriae.

Xenorhabdus spp., entomopathogenic bacteria symbiotically associated with nematodes of the family Steinernematidae, occur spontaneously in two phases. Only the phase I variants of Xenorhabdus nematophilus F1 expressed fimbriae when the bacteria were grown on a solid medium (nutrient agar; 24 and 48 h of growth). These appendages were purified and characterized. They were rigid, with a diameter of 6.4 (plusmn) 0.3 nm, and were composed of 16-kDa pilin subunits. The latter were synthesized and assembled during the first 24 h of growth. Phase II variants of X. nematophilus did not possess fimbriae and apparently did not synthesize pilin. Phase I variants of X. nematophilus have an agglutinating activity with sheep, rabbit, and human erythrocytes and with hemocytes of the insect Galleria mellonella. The purified fimbriae agglutinated sheep and rabbit erythrocytes. The hemagglutination by bacteria and purified fimbriae was mannose resistant and was inhibited by porcine gastric mucin and N-acetyl-lactosamine. The last sugar seems to be a specific inhibitor of hemagglutination by X. nematophilus.

Journal Article↗
PubMed1969

Antibody response of animals to Mycoplasma pneumoniae measured by latex agglutination.

The standardization, application, and usefulness of latex agglutination for Mycoplasma pneumoniae antibody titration were investigated and compared with tetrazolium reduction inhibition and complement fixation. The sera of guinea pigs and monkeys reacted in a specific fashion, whereas rabbit serum required pretreatment to eliminate its nonspecific agglutinin. Human serum given such pretreatment still contained a nonspecific agglutinin. The latex agglutination procedure compared favorably with complement fixation and metabolic inhibition tests for evaluating vaccine antigenicity.

Animals↗
PubMed2003

Rapid detection of contagious bovine pleuropneumonia by a Mycoplasma mycoides subsp. mycoides SC capsular polysaccharide-specific antigen detection latex agglutination test.

A latex agglutination test (LAT) has been developed for the diagnosis of contagious bovine pleuropneumonia (CBPP). The latex microspheres were coated with MmmSC polyclonal immunoglobulin G antiserum and detected MmmSC antigen in the serum of cattle infected with CBPP and in growth medium containing MmmSC. The specific antigen recognized by this test appeared to be the capsular polysaccharide (CPS). The LAT recognized all 23 strains of MmmSC examined in this study, with a sensitivity level of 2 ng of CPS, or the equivalent of 5 x 10(3) CFU, in a reaction volume of 0.03 ml. Therefore, rapid identification of MmmSC cultures should be possible. Agglutination was also observed with the related goat pathogens and "Mycoplasma mycoides" cluster members Mycoplasma mycoides subsp. mycoides large colony biotype (four of six strains positive) and Mycoplasma mycoides subsp. capri (three of six strains positive), in agreement with the suggestion that these latter two mycoplasmas may in fact represent a single species (although collectively exhibiting two capsular serotypes). Comparisons in diagnosis with the complement fixation test (CFT) were made by using African field sera from CBPP-infected cattle. After 2 (or 3) min of incubation, the test detected 55% (or 61%) of CFT-positive sera and 29% (or 40%) of CFT-negative sera, with an overall correlation in diagnosis of 62% (or 61%). The rates for false-positive diagnoses made by using "known" CBPP-negative sera from the United Kingdom were 3 or 13% after 2 or 3 min of incubation, respectively. The data agree with previous findings that some CBPP CFT-negative misdiagnoses may occur due to "antibody eclipsing" by excess circulating antigen. The LAT combines low cost and high specificity with ease of application in the field, without the need for any specialist training or equipment.

Animals↗
PubMed2002

Role of Fig2p in agglutination in Saccharomyces cerevisiae.

In W303-derived strains, disruption of FIG2 increased agglutinability of alpha cells, but not a cells, and did not alter expression of alpha-agglutinin, binding of 125I-labeled alpha-agglutinin, or mating efficiency. Fig2p overexpression led to alpha-cell-specfic suppression of agglutinability. These results imply that Fig2p is an indirect masker of the active sites in alpha-agglutinin.

Cell Adhesion↗
PubMed1970

Relationship of Serogroups of Neisseria meningitidis I. Microagglutination, Gel Diffusion, and Slide Agglutination Studies of Meningococcal Antisera Before and After Absorption with RAS-10 Strain of Meningococci.

Microagglutination tests were used to show the relationship of a nongroupable strain of Neisseria meningitidis (RAS-10) to other serological groups. RAS-10 antiserum has been prepared and studied for the first time. Antibodies to the RAS-10 strain were shown to be present in many grouping antisera obtained from different sources. These antibodies were absorbed from antisera to heterologous sero-groups with the RAS-10 strain. This procedure was shown to make antisera more specific by eliminating serological cross-reactions and false grouping of RAS-10 strains. Antisera before and after absorption with RAS-10 cells were studied by using double diffusion in gels. An antigen-antibody precipitation line for the RAS-10 meningococci was shown to be removed by this procedure. Antiserum to group 29E meningococci was absorbed with group Z cells, and precipitation lines for Z cells were removed. Group 29E antiserum agglutinated group 29E and group Z cells in the slide agglutination test but was specific for group 29E cells in this test after absorption with group Z cells.

Journal Article↗
PubMed1974

Ultrastructure of Mutants of Streptococcus mutans with Reference to Agglutination, Adhesion, and Extracellular Polysaccharide.

An electron microscopic study of S. mutans 6715-13 wild type and representatives of three distinct classes of glucan synthesis-defective mutants (which fail to form adherent microbial plaques but agglutinate normally in the presence of exogenous glucans) disclosed the presence of two sucrose-dependent, glucanase-sensitive, extracellular components. In the wild type, these extracellular glucans had predominantly fibrillar (with some globular) morphologies. However, in the mutant strains, there was a consistent reduction in or loss of the fibrillar components and dramatic increases in globular forms. A cell surface-associated fuzzy coat was consistently seen, and it was neither sucrose-dependent nor glucanase-sensitive. The data indicated that in vitro and in vivo adhesion and virulence at smooth tooth surfaces (all these properties dramatically reduced in the mutants) were causally and functionally related to the extracellular, fibrillar, glucan component, whereas in vitro glucan-mediated agglutination may be related to the cell-associated surface fuzzy coat.

Journal Article↗
PubMed1974

Agglutination of trypsinized sheep erythrocytes by the pili of Corynebacterium renale.

Corynebacterium renale strains which possessed numerous pili agglutinated trypsinized sheep erythrocytes, whereas those possessing only a few pili did not show such activity. By electron microscopy, the pili of C. renale adhered to the ghost of trypsinized sheep erythrocytes. After the treatment of C. renale with the homologous anti-pili serum, neither the agglutination of trypsinized sheep erythrocytes nor the adherence to the ghost by the pili were found. The adhesive property of the pili of C. renale was thus demonstrated.

Absorption↗
PubMed1984

Variability in expression of a cell surface determinant on Candida albicans as evidenced by an agglutinating monoclonal antibody.

The expression of a surface immunodeterminant of Candida albicans was investigated with an agglutinating immunoglobulin M monoclonal antibody. The 96 strains of C. albicans tested, of which 76% were recent clinical isolates, were capable of expressing the antigen. The antigen was also produced by strains of Candida tropicalis and Torulopsis glabrata, but not by other yeast species. Expression of the surface immunodeterminant in C. albicans varied as a function of growth as indicated by agglutinin reactions and indirect immunofluorescence tests. When yeast cells were tested with the agglutinin, three patterns of reactivity were observed. In general, cells in the early logarithmic phase were less reactive than cells in the mid-logarithmic phase. Antigen expression, as determined by agglutinin reactivity, was also influenced by the nutritional composition of the growth medium, and in general, cells from broth cultures were usually more reactive than cells grown on solid media. The antigen was solubilized from the cell surface of C. albicans by hot 1 M NaCl. These water-soluble extracts were capable of binding antibody, and a single precipitin band formed when soluble antigen was reacted with the monoclonal antibody in an Ouchterlony double-diffusion test. Whole cell preparations and hot NaCl extracts from yeast strains which did not agglutinate when mixed with the antibody also did not absorb the agglutinin from solution. These data indicate that the expression of surface antigens on C. albicans is a dynamic process which may be influenced by a number of environmental factors. The use of monoclonal antibodies may allow characterization of surface antigens presented to the host during candidiasis.

Absorption↗
PubMed1980

Agglutination of human erythrocytes by Fusobacterium nucleatum: factors influencing hemagglutination and some characteristics of the agglutinin.

Various aspects of agglutination of human erythrocytes by fusobacterium nucleatum were examined. Titration experiments done in buffered saline at pH 6 to 10 showed the same agglutination endpoint. The presence of high concentrations of NaCl in reaction mixtures did not alter titers, but KCl in concentrations of 0.5 to 3.6 M increased titers twofold. The agglutinin was inactivated by heat, acid, alkali, 5% Formalin, and the proteolytic enzyme subtilopeptidase A and therefore appeared to be a protein. Treatment of bacterial cells with 2-mercaptoethanol had no effect. Hemagglutination inhibition experiments revealed that arginine and other compounds containing a guanido group as part of their structure were inhibitory at low concentrations. Various hexoses, some hexose derivatives, and most common metal ions, when added to bacterium-erythrocyte mixtures, had no effect. The binding of dansyl-L-arginine to bacteria, but not erythrocytes, was demonstrable by ultraviolet fluorescence microscopy.

Agglutinins↗
PubMed1990

Isolation and characterization of mutants with lesions affecting pellicle formation and erythrocyte agglutination by type 1 piliated Escherichia coli.

The product of the pilE (also called fimH) gene is a minor component of type 1 pili in Escherichia coli. Mutants that have insertions in the pilE gene are fully piliated but unable to bind to and agglutinate guinea pig erythrocytes, a characteristic of wild-type type 1 piliated E. coli. In this paper we describe the isolation of 48 mutants with point lesions that map to the pilE gene. Such mutants were isolated by using mutT mutagenesis and an enrichment procedure devised to favor the growth of individuals that could form a pellicle in static broth containing alpha-methylmannoside, an inhibitor of erythrocyte binding and pellicle formation. Results indicated that the enrichment favored mutants expressing pilE gene products that were defective in mediating erythrocyte binding. Characterization of 12 of the mutants in greater detail revealed that certain lesions affected pilus number and length. In addition, a mutant that was temperature sensitive for erythrocyte binding was isolated and used to provide evidence that pellicle formation relies on the intercellular interaction of pilE gene products. Our results suggest a molecular explanation for the old and paradoxical observations connecting pellicle formation and erythrocyte agglutination by type 1 piliated E. coli.

Alleles↗
PubMed1980

Detection of Pneumococcal Capsular polysaccharide antigens by latex agglutination, counterimmunoelectrophoresis, and radioimmunoassay in middle ear exudates in acute otitis media.

The presence of pneumococcal antigen in middle ear exudates during acute otitis media was studied by latex agglutination and counterimmunoelectrophoresis. The positive antigen findings were confirmed by radioimmunoassay. Latex agglutination gave a positive result in 63% and counterimmunoelectrophoresis in 76% of samples that grew Streptococcus pneumoniae. The methods were complementary; the antigen was detected by one or both of the methods in 88% of these samples. Pneumococcal antigen was further detected in 15% of samples that grew other otitis pathogens and in 33% of samples in which no pathogenic bacteria were recovered by culture. The distribution of pneumococcal serotypes found by immunochemical methods only corresponded to that found by culture.

Child↗