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PubMed1991

Evaluation of bovine serum albumin preparations and Ficoll addition in capillary tube hemagglutination.

Eighteen commercial bovine serum albumin (BSA) preparations were evaluated for their efficacy in hemagglutination enhancement by the capillary tube method. Their physicochemical variables (pH, conductivity, free fatty acids, total protein, and polymer content) were quantified. The results showed that a BSA preparation which consistently performed well in hemagglutination enhancement contained some albumin polymer and had a low free fatty acid content. We also found that the addition of Ficoll (final concentration of 1.24%) could convert BSA preparations producing mediocre hemagglutination enhancement into good enhancers, provided that these preparations did not have high free fatty acid content. Hence, the addition of Ficoll can obviate the requirement for selecting a polymer-enhanced albumin for use in capillary tube tests.

Animals↗
PubMed1994

Correlation between cell-associated mannose-sensitive hemagglutination by Vibrio parahaemolyticus and adherence to a human colonic cell line Caco-2.

Cell-associated hemagglutination (cHA) activity with human erythrocytes was examined for 468 clinical and 71 environmental strains of Vibrio parahaemolyticus. Approximately 95% of the strains tested were cHA positive irrespective of source or Kanagawa phenomenon. 75% of clinical strains showed relatively strong mannose-sensitive hemagglutination (MSHA), whereas 88% of the environmental strains showed relatively weak mannose-resistant hemagglutination (MRHA). Adherence of V. parahaemolyticus to Caco-2 cells was also determined. A clear positive correlation between cell-associated MSHA and adherence to Caco-2 cells was observed.

Bacterial Adhesion↗
PubMed1969

Factors affecting the passive hemagglutination titration: dilution loops, titration trays, vibration, diluents.

Serial dilution with Takatsy loops resulted in exaggerated passive hemagglutination titers with most of the anti-bovine serum albumin sera tested. It appears that certain types of agglutinins adhere to the loop surface and are released only gradually. This adherence, or carry-over effect, was prevented by presoaking loops in gelatin or gelatin-rabbit serum-albumin solutions. Hemolysins did not adhere to loops. In general, hemagglutination reactions performed on plastic trays gave higher titers than those performed in glass test tubes. The quality of the hemagglutination pattern was dependent to a great extent on the type of plastic tray used. As much as a 100-fold difference in titers was obtained depending on the composition of the antiserum diluent. The increase in vibration, in terms of linear displacement (approximately twofold), resulted in an eightfold decrease in titers.

Animals↗
PubMed1973

Comparison of the direct agglutination and indirect hemagglutination tests in the determination of blood serum titers to Escherichia coli organisms.

A comparison of the direct agglutination test and the indirect hemagglutination test for the detection of blood serum antibodies to Escherichia coli organisms indicated that these serological tests were comparable. In some instances the indirect hemagglutination test provided higher endpoint readings. Preparation of the antigens for the indirect hemagglutination test was more time consuming than for the direct agglutination test. Crude extract and purified polysaccharides were comparable as red blood cell sensitizing agents.

Agglutination Tests↗
PubMed1974

Rubella hemagglutination inhibition: removal of nonspecific agglutination due to manganous chloride.

Nonspecific inhibitors of rubella hemagglutination can be removed by treatment of sera with heparin-manganous chloride for use in the hemagglutination-inhibition test. After removal of nonspecific inhibitors by this procedure, an excess of manganous chloride may remain. This may cause the cells to agglutinate, thus obscuring the reading at low serum dilutions. This disadvantage can be overcome by the addition of sodium carbonate, which forms an insoluble compound with manganous chloride and does not interfere with antibody determination. The procedure presents a further refinement of the hemagglutination inhibition test for rubella by increasing specificity and sensitivity; it permits detection of antibody levels as low as 1:4 in sera.

Agglutination↗
PubMed1978

Heterophil antigen in bovine sera detectable by immune adherence hemagglutination with infectious mononucleosis sera.

Bovine sera for tissue culture use were shown to contain heterophil antigen of the Paul-Bunnell-Davidsohn type by immune adherence hemagglutination tests with sera from patients with infectious mononucleosis. The antibody titers observed were comparable to those determined by two other methods; i.e., the immune adherence hemagglutination test was found to be as specific as the differential absorption test with horse erythrocytes or the ox cell hemolysis assay,but it appeared to be the most sensitive of the three procedures. All 16 individual or pooled bovine serum samples showed Paul-Bunnell-Davidsohn antigen at concentrations varying over a fourfold range. The potential usefulness of the immune adherence hemagglutination test for various Paul-Bunnell-Davidsohn antigen-related problems and implications of the observations are discussed.

Animals↗
PubMed1978

Association of hemolysin production, hemagglutination of human erythrocytes, and virulence for chicken embryos of extraintestinal Escherichia coli isolates.

One hundred forty-two strains of Escherichia coli isolated from extraintestinal infections were examined for colicin V (ColV) and hemolysin (Hly) production. For comparison, 20 strains isolated from the feces of normal individuals and 12 enteropathogenic strains of E. coli were tested for these properties. Thirty-five to 59% of extraintestinal isolates were Hly+, but only one fecal strain was Hly+. Colicin V biosynthesis was found for 12% of blood culture isolates, 7% of urine culture isolates and 16% of the strains from other extraintestinal infections. None of the fecal isolates was ColV+. Selected strains were tested for virulence in 13-day-old chicken embryos; these same strains were tested for their ability to hemagglutinate chicken or human erythrocytes. Of 22 extraintestinal isolates, 13 (59%) killed greater than or equal to 60% of the embryos within 72 h. Only one of eight normal fecal isolates and two of three enteropathogenic strains tested were virulent. About 80% of the virulent strains were Hly+. The most striking finding, however, was the hemagglutination of human erythrocytes by virulent extraintestinal isolates. It seems possible that the hemagglutination property reflects a specific common adherence factor.

Animals↗
PubMed1980

Serological differentiation between infected and vaccinated cattle by using purified soluble antigens from Brucella abortus in a hemagglutination system.

Serological tests used in current brucellosis eradication schemes, such as bacterial tube agglutination, do not readily distinguish between infected animals and those immunized with strain 19 or 45/20 Brucella abortus vaccines. In this study, sera from naturally infected cattle were used to identify serologically important antigens in extracts of virulent B. abortus by gel diffusion techniques. Antisera from rabbits hyperimmunized with selected precipitation lines were used for purification by affinity chromatography of two precipitating and one non-precipitating antigen from crude bacterial extracts. A passive hemagglutination test using these antigens was developed. A number of characterized bovine sera were screened by passive hemagglutination and conventional bacterial tube agglutination test. A considerable improvement in discrimination between sera from infected and vaccinated cattle was obtained with the hemagglutination test compared with bacterial tube agglutination.

Animals↗
PubMed1983

Hemagglutination and adhesiveness of epidemiologically distinct strains of Vibrio parahaemolyticus.

Twelve strains of Vibrio parahaemolyticus from four epidemiologically distinct groups were examined for their ability to hemagglutinate human, bovine, chicken, guinea pig, and rabbit erythrocytes and to adhere to human buccal mucosal epithelial cells in the presence and absence of mannose. Four of six Kanagawa-positive but none of six Kanagawa-negative strains showed mannose-sensitive hemagglutination with erythrocytes of rabbits and of one or more additional species. Mannose-resistant hemagglutination was shown by one strain in each group with no apparent relationship to strain source or hemolytic capability. All strains adhered to human buccal mucosal cells, with but a single strain showing significant difference in adherence at the alpha = 0.05 level. The adherence pattern had no relationship to the four epidemiological groups. Although adhesive processes may well be involved in disease caused by V. parahaemolyticus, our results do not support a role for adherence as a predictor of pathogenicity.

Adhesiveness↗
PubMed1989

Hemagglutinating properties of Shigella dysenteriae type 1 and other Shigella species.

Strains of Shigella dysenteriae type 1 cultured in Casamino Acids-yeast extract broth medium in the presence of 1 mM calcium chloride at 37 degrees C for 22 h induced hemagglutination of erythrocytes that was inhibited by N-acetylneuraminic acid, N-acetylneuramin-lactose, and alpha 1-glycoprotein. The hemagglutination was heat labile, and the absence of cell-surface appendages suggested a nonfimbrial adhesin(s). Under the same conditions, strains of Shigella flexneri (types 1a, 1b, 2a, and 2b) showed N-acetylneuraminic acid-resistant hemagglutination of erythrocytes.

Animals↗
PubMed1992

Epitope mapping of hemagglutinating adhesion HA-Ag2 of Bacteroides (Porphyromonas) gingivalis.

Thirteen monoclonal antibodies (MAbs) directed against hemagglutinating adhesion HA-Ag2 of Bacteroides (Porphyromonas) gingivalis were produced by immunizing mice with the relevant immunoprecipitate from crossed immunoelectrophoresis (CIE). Crossed immuno-affinoelectrophoresis and hemagglutination experiments confirmed that our MAbs recognized a molecule able to bind erythrocytes and involved in the hemagglutination process. In immunoelectron microscopy, these MAbs labelled amorphous material as novel cell-bound appendages distinct from fimbriae. CIE experiments allowed differentiation of the MAbs according to reactivity with immunoprecipitates Ag2, Ag8a, and Ag8c, which define HA-Ag2. The epitopes recognized by nine MAbs were mapped on three main antigenic domains (I, II, and III) by competition experiments and further grouped according to chemical composition and distribution on CIE immunoprecipitate. Domain I, defined by two MAbs, comprises an epitope with protein and carbohydrate determinants and distributed on Ag2 only. Epitopes of domain IIA, defined by four MAbs, are distributed on Ag8a, Ag8c, and Ag2 and are essentially composed of protein determinants but also have carbohydrate determinants that enhance the binding of the MAbs but are not essential. Epitopes of domain IIB, defined by two MAbs, and of domain III, defined by a single MAb, have a composition similar to that of domain IIA epitopes but are distributed on Ag8a and Ag8c only. A competition enzyme-linked immunosorbent assay with serum from normal subjects and patients with periodontitis suggested that domain I is more immunogenic than domain II.

Adhesins, Bacterial↗
PubMed1995

Hemagglutination and proteoglycan binding by the Lyme disease spirochete, Borrelia burgdorferi.

The ability of the Lyme disease spirochete to attach to host components may contribute to its ability to infect diverse tissues. We present evidence that the Lyme disease spirochete expresses a lectin activity that promotes agglutination of erythrocytes and bacterial attachment to glycosaminoglycans. Among a diverse collection of 21 strains of Lyme disease spirochete, hemagglutinating activity was easily detected in all but 3 strains, and these three strains were noninfectious. The ability to agglutinate erythrocytes was associated with the ability of the spirochete to bind to the sulfated polysaccharide dextran sulfate and to mammalian cells. Soluble dextran sulfate was a potent inhibitor of both hemagglutination and attachment to mammalian cells, while dextran had no effect on either activity, suggesting that dextran sulfate may inhibit attachment by mimicking host cell glycosaminoglycans. Consistent with this, the spirochete bound to immobilized heparin, and soluble heparin inhibited bacterial adhesion to mammalian cells. The bacterium did not bind efficiently to Vero cells treated with heparinase or heparitinase or to mutant CHO cell lines that are deficient in proteoglycan synthesis. Sulfation of glycosaminoglycans was critical for efficient bacterial recognition, as Vero cells treated with an inhibitor of sulfation, or a mutant CHO cell line that produces undersulfated heparan sulfate, did not mediate maximal spirochetal binding. Binding of the spirochete to extracellular matrix also appeared to be dependent upon this attachment pathway. These findings suggest that a glycosaminoglycan-binding activity which can be detected by hemagglutination contributes to the attachment of the Lyme disease spirochete to host cells and matrix.

Animals↗
PubMed1966

Serological studies of types A, B, and E botulinal toxins by passive hemagglutination and bentonite flocculation.

Johnson, H. M. (Robert A. Taft Sanitary Engineering Center, Cincinnati, Ohio), K. Brenner, R. Angelotti, and H. E. Hall. Serological studies of types A, B, and E botulinal toxins by passive hemagglutination and bentonite flocculation. J. Bacteriol. 91:967-974. 1966.-Formalinized sheep red blood cells (SRBC), sensitized with types A, B, and E botulinal toxoids and toxins by bis-diazotized benzidine (BDB), were tested against A, B, and E antitoxins prepared in horses and rabbits. Type B antitoxin cross-reacted with A toxoid SRBC, but the reciprocal cross-reaction was not observed. E toxin SRBC were specifically agglutinated by E antitoxin. Flocculation of antigen-sensitized bentonite particles was less sensitive in titration of antitoxin than hemagglutination. Also, reciprocal cross-reactions were observed between types A and B antitoxins. Cross-reactions in both serological tests were eliminated by titration of antitoxins in the presence of the heterologous antigens, with no inhibitory effect on the homologous antitoxins. Generally, equine antitoxins were less suitable for agglutinations, especially of antigen-sensitized bentonite particles. Types A, B, and E antitoxins were specifically inhibited by 43, 39, and 245 mouse ld(50) of their respective homologous toxins in the hemagglutination-inhibition test. A, B, and E antitoxins were specifically inhibited by 500, 950, and 1,500 mouse ld(50) of their respective homologous toxins in bentonite flocculation inhibitions. Formalinized SRBC sensitized with rabbit types A and B antitoxins by BDB were respectively clumped by as little as 0.75 to 1.3 mouse ld(50) of A toxin and 2.3 ld(50) of B toxin, whereas bentonite particles sensitized by the same antitoxins were specifically clumped by 150 ld(50) of A toxin and 630 ld(50) of B toxin. E antitoxin sensitization of SRBC or bentonite particles was not successful. Evidence is presented that indicates that the serological procedures are applicable to the detection of botulinal toxins in food.

Animals↗
PubMed1966

Identification of enteroviruses by hemagglutination-inhibition.

Kern, Jerome (Pacific Research Section, Honolulu, Hawaii), and Leon Rosen. Identification of enteroviruses by hemagglutination-inhibition. J. Bacteriol. 91:1936-1942. 1966.-Approximately 40% of a group of 906 enterovirus isolates cytopathic for monkey cell cultures were found to possess hemagglutinins for human erythrocytes when tested at temperatures of 4 and 37 C and at pH 5.8 and 7.3. The hemagglutinating isolates could be classified by relatively simple techniques into 18 serotypes. Four of these serotypes, echovirus type 24 and coxsackievirus B types 1, 5, and 6, had not previously been known to include hemagglutinating strains. One serotype, Toluca-3, represented a previously unrecognized enterovirus, and two other serotypes may also represent previously unrecognized enteroviruses.

Enterovirus↗
PubMed1979

Indirect hemagglutination test for detection of antibody to Rickettsia rickettsii in sera from humans and common laboratory animals.

Antibody production in humans and three species of laboratory animals infected with Rickettsia rickettsii was determined with the indirect hemagglutination test. Rabbits, guinea pigs, and mice were inoculated with R. rickettsii and bled at intervals. Antibody which agglutinated both fresh and glutaraldehyde-fixed sheep erythrocytes sensitized with antigen prepared either from purified rickettsiae or from infected yolk sacs was found in rabbit sera at all intervals tested (10 to 59 days postinfection). Antibody which agglutinated fresh but not glutaraldehyde-fixed erythrocytes sensitized with either of the above antigens was detected in guinea pig sera obtained 7, 14, and 28 days postinfection. Antibody was found in mice inoculated with 5.6 x 10(6) plaque-forming units of R. rickettsii but not in mice given 5.6 x 10(2) plaque-forming units. Peak indirect hemagglutination titers occurred in nonvaccinated human Rocky Mountain spotted fever patients about 3 weeks after onset of illness, and antibody was still detectable after 1 year. Both human immunoglobulin G and human immunoglobulin M antibodies agglutinated sensitized cells, but immunoglobulin M antibodies apparently were more efficient. The indirect hemagglutination test is useful for the titration of human, rabbit, guinea pig, and mouse antibodies when the appropriate erythrocytes are used.

Animals↗
PubMed1979

Analysis of Vibrio parahaemolyticus soluble antigens by employing passive hemagglutination.

The passive hemagglutination assay was explored as a sensitive test of immunological reactivity of endotoxin and other antigens prepared from selected strains of Vibrio parahaemolyticus. The tannic acid procedure for passive hemagglutination, commonly used with protein antigens, was the only procedure yielding good results with V. parahaemolyticus protein extracts, endotoxins, and related preparations. These results were probably due to the presence of large amounts of protein in the V. parahaemolyticus endotoxins as determined by earlier work referenced in the text. Glucose and galactose as possible antigenic determinants in the endotoxin of a Vibrio strain were tested by inhibition tests. Cross-reactions were observed between endotoxin preparations, but were low in hemagglutination, suggesting reactions of common generic antigens. The ability of V. parahaemolyticus endotoxins to stimulate production of antibodies was determined.

Antibodies, Bacterial↗
PubMed1981

Nonspecific reactions in the hemagglutination inhibition test for detection of rubella antibodies.

Approximately 7% of the sera tested to determine the presence of rubella-specific antibodies by the hemagglutination inhibition test demonstrated abnormal patterns of reactivity, rendering the test unreadable. Another 3% of sera were shown to have false-positive titers as high as 1:128. When these abnormally reacting and false-positive sera were heated at 56 degrees C for 30 min after chemical treatment they always converted to negative, indicating the absence of specific rubella hemagglutination-inhibiting antibody. These results were confirmed by fractionation of the sera after sucrose gradient centrifugation. It was established that manifestation of these nonspecific results was dependent on the concentration of Ca2+ or Mn2+. The heat-labile inhibitor(s) responsible for abnormal and false-positive reactions was found not to be complement. This inhibitor(s) was detected in the light fractions of sera and when added to negative sera was capable of reproducing the abnormal patterns of reactivity. These results emphasize the necessity of heating sera for the rubella hemagglutination inhibition test after the chemical removal of nonspecific inhibitors.

Antibodies, Viral↗
PubMed1981

Freeze-dried erythrocytes for an indirect hemagglutination test for detection of cytomegalovirus antibodies.

An indirect hemagglutination test with lyophilized, fixed, tanned, and cytomegalovirus (CMV)-sensitized sheep erythrocytes for the detection of CMV antibodies is reported. To avoid nonspecific hemagglutination, cells were fixed with glutaraldehyde or Formalin directly in whole blood. The lyophilized, CMV-sensitized erythrocytes obtained by this technique were stable up to 9 months at 37 degrees C and retained the same reactivity at fresh, CMV-sensitized cells. Indirect hemagglutination performed with lyophilized, sensitized cells was highly efficient in detecting CMV-antibodies as compared with complement fixation and enzyme immunoassay.

Animals↗