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Early detection of streptococcal group antigens in skin samples by latex particle agglutination.

The streptococcal group antigens (A to G) were researched in skin biopsy specimens by a latex agglutination technique using commercial (Slidex Streptokit bio-Mérieux and Streptex Wellcome) and experimental reagents. Twenty-seven patients with obvious cutaneous infections (12 cases of erysipelas, nine cases of cellulitis, and six cases of necrotizing fasciitis) and 27 age- and sex-matched controls were studied. Our preliminary data demonstrated the possibility of an early diagnosis of streptococcal cutaneous infections involving the deep dermis and/or hypodermis. We used a latex agglutination technique that is quite specific and gives a better sensitivity (0.63) than either conventional bacteriologic cultures on needle aspiration or skin biopsy specimens.

Adult↗

[Serology of Mediterranean boutonneuse fever. Kinetics of antibodies detected by 3 methods: indirect immunofluorescence, indirect hemagglutination and latex agglutination].

The purpose of this work was to determine the kinetics of antibodies in mediterranean spotted fever as determined by three serologic methods: indirect fluorescent antibody test, latex agglutination test and indirect hemagglutination test. No difference was noticed in the early kinetics but after 6 months, the latex agglutination test and the indirect hemagglutination test did not detect antibodies but the indirect fluorescent antibody test was still positive. This reaction is convenient for seroepidemiologic studies.

Antibodies, Bacterial↗

[Hemobacterial agglutination--method of determining antierythrocyte antibodies].

A technique of antierythrocyte antibody detection is described. It as based on co-agglutination of red blood cells and S. aureus cells. The method is unsophisticated and non-laborious. It provides massive agglutination, its sensitivity is one order of magnitude higher than that of the Coombs test.

Anemia, Hemolytic, Autoimmune↗

A study of complement-fixation and serum agglutination tests on vibrio fetus infected and immunized cattle.

Complement-fixation (CF) and tube agglutination (TA) tests for demonstration of Vibrio fetus antibodies were conducted on the sera of three groups of ten heifers. One group was vaccinated subcutaneously with a commercial V. fetus var venerealis bacterin and challenged intra-utero, at the external os cervicus one month later; the second was infected only and the third vaccinated only. The vaccinated cattle developed high CF serum titers, but no such increase was observed in animals infected only. A moderate increase in serum antibody titers was demonstrated by the TA test following either infection or vaccination; although titers observed were not higher than those observed in the sera of some apparently normal uninfected animals. The group receiving both vaccine and challenge was the only one in which significant serum antibody titers were demonstrable by the TA test. The sera of these animals also had significant titers in the CF tests. The CF and TA tests detected serum antibodies produced by the parenteral inoculation of V. fetus antigen. These two tests were of limited value in detecting serum antibodies from animals with genital V. fetus var venerealis infection, although the formation of local antibodies was demonstrable by the vaginal mucus agglutination test.

Animals↗

A routine method for assaying the anti-toxoplasma IgM antibodies. Latex agglutination test on human sera preabsorbed with protein A-bearing staphylococcal cells.

The authors investigated the procedure which demonstrates, by means of the latex agglutination test (LA) the residual IgM antibody activities in sera preabsorbed with protein A-bearing Staphylococcus aureus and evaluated the usefulness of this technique with the following results. (1) LA (Toxotest-LA, Eikenkagaku Co.) gave clear-cut and reproducible agglutination patterns. Its specificity was comparable to that of HA and its sensitivity was somewhat lower than that of HA. (2) The absorption of 0.2 ml of 1:10 dilution of many test sera with the sediment of 0.5 ml of a 10% bacterial suspension for ten minutes or more provides the suitable condition for the absorption procedure. (3) HA-positive 183 sera revealed no residual antibody activities after absorption. These are supposed to derive from persons with a long-standing infection. Sera from 4 laboratory infection cases, from 2 toxoplasmic lymphadenopathy cases and from a pregnant woman, showed more or less residual activities. In 5 out of 7 sera tested, the residual activities were 2-ME sensitive, indicating the presence of specific IgM antibodies. (4) The specific and the total IgM were reduced slightly by the absorption procedure.

Absorption↗

Comparison of indirect double antibody and double antibody sandwich ELISA techniques with latex agglutination test for the diagnosis of human rotavirus infection.

94 faecal samples from infants and children suffering of acute gastroenteritis were investigated for rotavirus by indirect double antibody sandwich ELISA kit (WHO, Geneva), Rotavirus ELISA kit (DAKOPATTS A/S, Copenhagen) and Rotalex latex-agglutination kit (Orion Diagnostica, Helsinki). The ELISA techniques gave almost identical results and seemed to be of same sensitivity and specificity. Rotalex agglutination had an overall agreement of 88% with ELISA. It is concluded that strongly positive reactions found by Rotalex may be regarded as true positive reactions, whereas samples producing weakly positive and/or negative reactions should be retested in a more specific and sensitive assay, such as enzyme linked immunosorbent-assay (ELISA).

Enzyme-Linked Immunosorbent Assay↗

Precipitating and agglutinating activity in duck anti-soluble protein immune sera.

Ducks were inoculated parenterally, up to 4 times, with bovine serum albumin (BSA), B gamma globulin (BGG), or horse gamma globulin (HGG). Their sera were tested in immunodiffusion tests for precipitins. Only BSA and BGG induced precipitins. Only 7 of 443 sera tested, obtained from 3 of 62 inoculated ducks, developed a precipitin line with homologous antigen. All 7 sera were obtained from the earliest bleeding (7 days) after inoculation. Sera were also tested for agglutinins in direct passive hemagglutination (DPHA) and direct red-cell-linked antigen tests (DRCLAT) and for nonagglutinating antibodies in indirect red-cell-linked antigen tests (IRCLAT). No duck had a passive-hemagglutination-demonstrable primary immune response. Demonstrable DPHA titers after subsequent inoculations were very low compared with responses to these antigens of other species noted by other workers. Duck immune response was greatly diversified: all inoculation regimens that induced agglutinins in some ducks left others completely unstimulated. Precipitins and agglutinins of the same ducks correlated well: those 7 sera that had precipitins also had the highest DPHA titers for the 3 donor ducks. However, hemagglutinating titer and presence of precipitins in sera of different ducks correlated poorly. Increasing the age of the ducks at the first inoculation from 6 to 10 weeks increased the number of responders and DPHA titers of their sera. Non-agglutinating antibodies were demonstrated in anti-BSA sera: in IRCLAT, the sera had titers 2-to-256-fold higher than the sera in DPHA or DRCLAT had. Duck immunoglobulins were deficient in those immunological reactions (precipitation and agglutination) that require functional bivalency.

Animals↗

Cell-cell recognition in yeast. Characterization of the sexual agglutination factors from Saccharomyces kluyveri.

The cell surface molecules responsible for sexual agglutination between haploid cells of opposite mating type from Saccharomyces kluyveri have been purified and characterized. The 17-factor, released from 17-cells by beta-glucanase digestion (Zymolyase), is a glycoprotein of 6 X 10(4) Da. Its binding activity is heat- and protease-labile, but it is stable to reducing agents and exo-alpha-mannosidase digestion. The 16-factor, released from 16-cells by Zymolyase digestion, has a molecular weight of 5 X 10(5) and is over 95% carbohydrate. An active binding fragment can be released from 16-factor, from the factor purified from a mutant of 16-cells (16(mnn1)-factor), and from the surfaces of the cells themselves by dithiothreitol treatment. The 16(mnn1)-binding fragment has a molecular weight of 2 X 10(4) and is 30% carbohydrate. Its binding activity is stable to heat and some proteases, but it is labile to pronase, carboxypeptidases A and Y, alpha-mannosidases, and mild periodate treatment. 125I-16(mnn1)-binding fragment adheres specifically to 17-cells but does not bind to 16-cells or cells of other yeast strains. The binding of the labeled fragment to 17-cells is characterized by a KA of 10(8) M-1, and 5 X 10(5) binding sites are present per cell. The purified intact factors are monovalent and appear to interact in a lock and key fashion to cause the specific agglutination of S. kluyveri 16- and 17-cells.

Amino Acids↗

["Sugar-agglutinability"].

Haemagglutination induced by saccharose is shown to go parallel with a binding of autologous isohaemagglutinin to the surface of erythrocytes. Under suitable conditions, i.e. in the presence of anorganic ions and health, haemagglutinin may be largely separated from the cell surface and the saccharose agglutination may be inhibited. Haemagglutination induced by saccharose will re-appear after agglutinins being re-adsorbed at low incubation temperatures and prolonged incubation time in non-ional as well as ional media. Therefore it is assumed that in the inert non-ional atmosphere the autologous haemagglutinin will produce a phenomenon of agglutination in non-homologous erythrocytes similar to that in homologous cells.

Erythrocytes↗

The molecular mechanism of cryoprecipitation and cold agglutination of an IgM lambda Waldenström macroglobulin with anti-Gd specificity: sedimentation analysis and localization of interacting sites.

The physicochemical properties and immunologic specificity of an IgM lambda cryoglobulin with cold agglutinin activity (designated MAT) were investigated. Discrete binding sites involved in cryoprecipitation were shown to exist on the reductive 7S subunits and tryptic Fab mu and Fc mu 5 fragments derived from MAT. Analytical ultracentrifugation indicated the nonprecipitating MAT subunits self-associated into rapidly sedimenting paucidisperse boundaries of 32S, 25S, and 8.7S at 13 degrees C, which were converted to a monodisperse 5.7S boundary at 35 degrees C. Furthermore, MAT Fab mu and Fc mu 5 fragments exhibited association with each other at 13 degrees C but not at 35 degrees C when studied by analytical ultracentrifugation. The cold agglutinin activity of MAT can be inhibited by sialyllactose or treatment of erythrocytes with neuraminidase but not papain. Therefore, the membrane determinants of human erythrocytes are Gd (glycolipid-dependent or protease-resistant glycoproteins) with terminal sialic acid. Treatment of MAT with neuraminidase abolished its precipitating properties, but had no effect on its ability to agglutinate untreated erythrocytes. The cryoprecipitation and cold agglutination phenomena exhibited by MAT therefore seem to involve identical or cross-reacting carbohydrate antigens with terminal sialic acid residues.

Agglutinins↗

Interaction of platelets, von Willebrand factor, and ristocetin during platelet agglutination.

Ristocetin induces platelet agglutination in the presence of human factor VIII-associated ristocetin cofactor (vWF). The specificity, extent, and tenacity of binding among these reactants during agglutination and deagglutination were examined. Purified human vWF polymers were radioiodinated and reisolated. Radioiodinated vWF, a disulfide-linked polymer of 230,000 dalton subunits, attached to formalinized human platelets only in the presence of ristocetin. This binding reached equilibrium within 30 sec, and as ristocetin concentrations were raised from 0.2 mg/ml, the extent of attachment increased progressively to reach maximum at 0.5 to 0.6 mg/ml ristocetin. Ristocetin-induced binding was inhibited by vancomycin, unlabeled-purified vWF polymers, normal and hemophilia A plasma, and rabbit anti-human vWF. Binding was not impaired by plasma without detectable vWF or by naturally occurring human IgG antibodies to factor VIII coagulant activity. When formalinized platelets were pelleted from suspensions containing 125I-ristocetin, small quantities of radiolabeled ristocetin associated with platelets both in the presence or absence of vWF. About 95% of the attached 125I-ristocetin was removed by subsequent washes in buffered saline. The attachment of unmodified ristocetin or 125I-ristocetin to platelets, or the formation of complexes with vWF, could not be detected by agarose column chromatography, sucrose cushion ultracentrifugation, or equilibrium dialysis. These results indicate that (1) the initial binding of human vWF polymers to platelets is a specific interaction which requires the presence of ristocetin; (2) ristocetin and human vWF do not form persistent complexes in solution; and (3) the association of ristocetin and platelets is of low affinity.

Blood Coagulation Factors↗

[The latex agglutination test for the diagnosis of meningococcal A and C, pneumococcal and H. influenzae b meningitis. A study of 920 purulent cerebrospinal fluids (author's transl)].

The cerebrospinal fluids of 920 Senegalese patients with purulent meningitis wee examined by latex particle agglutination (LA) and the results were compared with those of conventional bacteriology and of counter immuno electro phoresis (C.I.E.). For all three organisms taken globally, CSF culture was positive in 72.7% of the patients, the LA test in 78.8% and the C.I.E. test in 89.0%. The LA test was positive in 17.9% of subjects with negative CSF culture. When the results wee analysed for each organism separately, the LA test was positive in 69.3%, 74.7% and 85.9% respectively of patients with meningococcal, pneumococcal and H. influenzae b meningitis. False agglutinations were found in 6.7% of meningitis due to other pathogens. Cross-reactions and autoagglutinations were rare (0.4% and 3.7% respectively). The sensitivity, specificity, simplicity and rapidity of the LA test make it extremely useful for the diagnosis of purulent meningitis, particularly in Africa.

Haemophilus influenzae↗

Influence of agglutinating antibody in experimental cryptococcal meningitis.

A model for chronic Cryptococcus neoformans meningitis in corticosteroid-treated rabbits was used to determine the influence of pre-formed agglutinating antibody to cryptococcal polysaccharide on the progress of this infection. Immunized rabbits developed serum agglutinating antibody with a geometric mean titre of 1:32, but none was detected in cerebrospinal fluid. Prior immunization did not enhance immunity to infection, had no effect on the number of viable cryptococci in cerebrospinal fluid, and did not prevent dissemination outside the central nervous system. Future investigations in this field should focus on cellular rather than humoral defence mechanisms.

Agglutinins↗

[A serological study of the contagious equine metritis: comparison between indirect immunofluorescence, slow agglutination and complement fixation techniques (author's transl)].

Serological response of pony mares to contagious equine metritis is studied comparing three techniques: slow agglutination, complement fixation and indirect immunofluorescence. Sera were taken from pony mares vaccinated with a heat inactivated suspension of Haemophilus equigenitalis, from experimentally-infected pony mares and from healthy horses. All three reactions detected antibodies in vaccinated and infected animals. The highest titers are observed with vaccinated mares. Titers are low in infected animals. Antibodies detected by indirect immunofluorescence appeared sooner and persisted longer in diseased animals than agglutinating or complement fixing antibodies. Only indirect immunofluorescence revealed a new contamination of two mares following coitus with a stallion excreting H. equigenitalis. Indirect immunofluorescence must be recommended in diagnosis of contagious equine metritis and in detection of chronic carriers.

Animals↗

Detection of amino acids as possible promoters of bladder cancer in rats by measuring their enhancement of agglutination of bladder cells by concanavalin A.

The effects of amino acids on the enhanced agglutinability of bladder cells with concanavalin A induced by subcarcinogenic treatment with N-butyl-N-(4-hydroxybutyl)nitrosamine were examined. The amino acids examined were L-alanine, L-arginine, L-asparagine, L-aspartic acid, L-cysteine, L-glutamic acid, L-glutamine, L-glycine, DL- and L-histidine, L-hydroxyproline, L-isoleucine, D- and L-leucine, L-lysine, L-methionine, DL- and L-phenylalanine, L-proline, L-serine, L-threonine, DL-, D- and L-tryptophan, L-tyrosine and D- and L-valine. They were added to powdered diet at a concentration of 2.0%. L-Leucine, L-isoleucine, L-valine, DL- and D-tryptophan prolonged the period during which the bladder cells showed enhanced agglutinability with concanavalin A. Leupeptin, a protease inhibitor, and L-leucyl-L-leucine were also examined at a concentration of 0.1% because of their similar chemical structures, and were found to have the same effect. The tumor-promoting effects of DL-tryptophan and leupeptin have already been established by in vivo carcinogenesis experiments. The effects of L-leucine, L-isoleucine, L-valine, D-tryptophan and L-leucyl-L-leucine, detected by this short term assay, suggest that these compounds may also be promoters of bladder cancer in rats.

Amino Acids↗

[Serodiagnosis of pregnancy in the cow with the latex passive agglutination technic].

This paper describes a test for diagnosing pregnancy in the cow based on the detection of blastocyst antigens in the maternal blood circulation. Rabbit anti-bovine embryo serum was agglutinated with the sera of non-pregnant cows to obtain specific antibodies. These rabbit antibodies were then absorbed on latex beads which could then be used for the standard passive agglutination reaction. This diagnostic technique was tested on 415 animals. The results on pregnant cows were successful in 78 p. 100 of the cases (confidence interval: 69-86 p. 100) and on non-pregnant cows 94 p. 100 of the time (confidence interval: 90-97 p. 100). This test is independent of the status of the ovaries and of the pregnancy stage. Detection is reliable from day 26 after coitum.

Animals↗

[Mechanism of the erythrocyte de-agglutination reaction].

The effect on the indication sensitivity of the level of antigenic sensitization of erythrocytes, molecular parameters of sensitin and of the antigen determined was revealed on various models of agglutinates and antigens determined with the aid of erythrocyte deaggregation reaction (EDR). By its dependence on the EDR sensitivity from the hemosensitization level this reaction was analogous to the passive hemagglutination inhibition test described earlier. Specific deaggregation activity of the antigens depended on their molecular parameters in the same way as their specific agglutinating activity in respect to the erythrocytic antibody diagnostic agents--both of these properties were more intensive in larger antigenic molecules.

Agglutinins↗