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PubMed1979

Rapid diagnosis of type III group B streptococcal meningitis by latex particle agglutination.

A latex particle agglutination assay was developed to detect type III group B streptococcal antigen in cerebrospinal fluid. Agglutination was observed in each of 12 initial and 54% of 26 subsequent CSF specimens obtained a mean of 53 hours after the institution of antimicrobial therapy. Latex agglutination was more sensitive than countercurrent immunoelectrophoresis for the detection of type III group B streptococcal antigen in CSF from neonates and young infants with meningitis.

Antigens, Bacterial↗
PubMed2003

Large-scale validation of a latex agglutination test for diagnosis of tuberculosis.

Large-scale validation of a simple latex agglutination test for the diagnosis of tuberculosis is described. Soluble antigens extracted from a non-pathogenic saprophytic mycobacterium, Mycobacterium w, which shares antigenic determinants with Mycobacterium tuberculosis, were covalently linked to carboxylated polystyrene latex beads. Batch to batch reproducibility of coated latex was ensured. Latex reagents were standardized to overcome non-specific agglutination. Reagents of the test are stable for 1 year at 4 degrees C. A total of 1,058 serum samples of pulmonary and extrapulmonary tuberculosis patients or patients with other pulmonary diseases and healthy controls living in endemic areas were tested. Sensitivity of 94% for pulmonary tuberculosis and 87% for extrapulmonary tuberculosis was obtained. Specificity is 92.2% for healthy controls and patients with other respiratory diseases. We conclude that the latex agglutination test can be utilized for mass screening for both pulmonary and extrapulmonary tuberculosis where diagnosis by existing methods is much more difficult.

Adolescent↗
PubMed1990

Monoclonal antibody-coated latex agglutination assay for identification of Actinobacillus actinomycetemcomitans.

Three monoclonal antibodies (MAbs) to lipopolysaccharide of Actinobacillus actinomycetemcomitans strain Y4 (serotype b) and eight MAbs to a serotype b-specific polysaccharide antigen of strain Y4 were obtained. Latex particles sensitized with an MAb to the Y4 lipopolysaccharide produced a positive agglutination with whole cells of all three serotypes of A. actinomycetemcomitans, but not with Haemophilus aphrophilus, Haemophilus paraphrophilus, Haemophilus influenzae, Porphyromonas (Bacteroides) gingivalis, "Bacteroides" intermedius, Fusobacterium nucleatum and Escherichia coli. On the other hand, latex particles sensitized with an MAb to the serotype b-specific polysaccharide antigen agglutinated with whole cells of serotype b A. actinomycetemcomitans and P. gingivalis, but not with heated and trypsinized cells of P. gingivalis. The simple and rapid latex agglutination assay using MAbs may be useful for the identification of A. actinomycetemcomitans.

Actinobacillus↗
PubMed2002

[Validity of pretransfusion bedside agglutination tests for pretransfusional control].

The incidence rate of ABO hazards of transfusion remains high in France. In this country, bedside pretransfusion controls include an agglutination test for red cells only, although its validity has scarcely been assessed in the practice. 847 nurses from 9 public hospitals and private clinics in a French region participated in a study aimed at measuring the sensitivity and specificity of pretransfusion bedside agglutination tests within hospital wards. Sensitivity was found to be 93.9% +/- 3%. Nondetection of mismatching was increased by two risk factors only: having worked more than 4 years in the same ward, and not having been trained to use this test. The sensitivity of this test might still be improved. This test is found sensitive enough to be kept. Nevertheless, if used alone, it is not a safe protection against recipient's mismatch. Authors recommend both to improve agglutination test sensitivity and to link it strongly to the bedside checking of both transfusion information and the recipient's identity.

ABO Blood-Group System↗
PubMed1994

[Measurement of immunologic agglutination of erythrocytes by ultrasonic interferometry].

Ultrasound interferometry is a new methodology which has been developed in our laboratories in order to measure precisely and quickly the size of particles sedimenting in liquid on horizontal surface, upon gravity. Applied to red blood cells, this method evaluates the sedimentation of erythrocytes, their aggregation induced by proteins or aggregating compounds as well as their agglutination upon immune reactions. The quantitative assessment of red cell agglutination was applied to the study of blood groups and to the search for red cell antibodies. Preliminary results show that ultrasound interferometry is 1) quantitative, measuring the size of agglutinates; 2) sensitive; 3) specific; 4) fast; 5) able to detect irregular antibodies.

Erythrocytes↗
PubMed1997

[Rheologic study of the dissociation reaction of erythrocyte immunologic agglutinates: application to subgroup A erythrocytes].

Red blood cell Agglutinates can be dissociated by providing enough energy which may be supplied by shear stress in a viscometer. In this paper a new technique is proposed to determine dissociation parameters of erythrocyte immunological agglutinates by using a laser backscattering method. Backscattered light intensity increases during the process of agglutinate dissociation when a controlled shear stress in a transparent Couette viscometer is applied. The obtained curves of dissociation can be fitted by an exponential function. A numerical integration of the dissociation curves allows us to obtain a parameter associated to the energy used in the dissociation process. A satisfactory differentiation of erythrocyte sub-groups Al, Az, A,B and Am has been carried out using this technique.

Blood Viscosity↗
PubMed2003

Macrocyclic glycoclusters. Self-aggregation and phosphate-induced agglutination behaviors of calix[4]resorcarene-based quadruple-chain amphiphiles with a huge oligosaccharide pool.

Macrocyclic glycocluster compounds 2n (n = 2-7) with four alkyl (undecyl) chains and eight oligosaccharide moieties on the opposite sides of the calix[4]resorcarene macrocycle are prepared from the reactions of the corresponding octaamine derivative with maltooligosaccharide lactones. Combined evidence from dynamic light scattering (DLS), gel permeation chromatography (GPC), and transmission electron microscopy (TEM) indicates that they form small micelle-like nanoparticles (d congruent with 3 nm) in water. In the presence of Na2HPO4/NaH2PO4, nanoparticles are agglutinated with phosphate ions as a glue to grow in size up to 60-100 nm, as revealed by DLS as well as microscopy (TEM and AFM). The phosphate-induced agglutination processes can be followed by surface plasmon resonance (SPR). Amphiphile 2n is readily immobilized on the hydrophobized sensor chip of SPR to give a closely packed monolayer with oligosaccharide moieties exposed to bulk water. While there is no further adsorption of 2n on the resulting monolayer, this does occur when the latter is pretreated with the phosphate salts, ultimately giving rise to a multilayer upon repeated treatment of the chip with 2n and Na2HPO4/NaH2PO4 in an alternate manner. Kinetic analyses show that the phosphate-mediated inter(saccharide) interactions in terms of rate and affinity are markedly dependent on the oligosaccharide chain lengths (n), becoming more favorable with increasing n's. The novel aggregation and agglutination behaviors observed are discussed in terms of immobilizable and irreversible micelles on the basis of the cone-shaped structure of quadruple-chain amphiphile 2n having a huge saccharide pool and the efficiency of multiple hydrogen bonding therein. The unique intermolecular binding properties of compound 22 and analogues so far reported are reviewed in light of the present finding.

Bridged-Ring Compounds↗
PubMed1974

Navy (Haricot)-bean (Phaseolus vulgaris) lectin. Isolation and characterization of tow components from a toxic agglutinating extract.

Two protein components were isolated in a highly purified state from a toxic fraction from the navy (haricot) bean (Phaseolus vulgaris). One, a lectin, strongly agglutinated horse erythrocytes and leucocytes, agglutination being readily observable with both types of cell at a lectin concentration of 4mug/ml. The other component (component 1), although possessing some similarity in composition, was thought to be non-agglutinating or, at most, only very weakly so. Component 1 had a mol.wt. of about 143000 and a subunit mol.wt. of about 37000, suggesting a tetrameric structure probably with identical subunits. Alanine was the only N-terminal amino acid identified and the molecule was notable in being devoid of tryptophan and cysteine, low in methionine and high in leucine, glutamic acid and aspartic acid. The lectin was somewhat smaller (mol.wt. about 114000) and apparently also composed of four identical subunits of mol.wt. about 30000. Dansylation showed that arginine occupied the N-terminus of the polypeptide chain. Aspartic acid, serine, threonine and leucine were the predominant amino acids of the lectin, and the sulphur-containing amino acids were entirely absent. Both constituents were glycoproteins and the compositions of the carbohydrate portions (4.9% for component 1 and 8.1% for the lectin) were generally similar, consisting of mannose and glucosamine with smaller amounts of glucose and traces of xylose and arabinose.

Alanine↗
PubMed2004

Crystal structure of a platelet-agglutinating factor isolated from the venom of Taiwan habu (Trimeresurus mucrosquamatus).

Platelet glycoprotein Ib (GPIb)-binding proteins (GPIb-BPs) from snake venoms are usually C-type lectins, which target specific sites of GPIbalpha and elicit distinct effects on platelets. In the present paper, we report a tetrameric platelet-agglutinating factor (molecular mass 121.1 kDa), termed mucrocetin, purified from the venom of Taiwan habu (Trimeresurus mucrosquamatus ). Mucrocetin is a GPIbalpha agonist with a binding site distinct from that of flavocetin-A (a snake venom GPIbalpha antagonist) on GPIbalpha, in spite of the high sequence identity (94.6%) between the two venom lectins. The crystal structure of mucrocetin was solved and refined to 2.8 A (1 A=0.1 nm) resolution, which shows an interesting crystal packing of six-layer cylinders of doughnut-shaped molecules. The four alphabeta heterodimers are arranged in an unusual square-shaped ring stabilized by four interdimer 'head-to-tail' disulphide bridges. Detailed structural comparison between mucrocetin and flavocetin-A suggests that their disparate platelet effects are probably attributable to different charge distributions on the putative concave binding surface. A unique positively charged patch on the binding surface of mucrocetin, formed by Lys102, Lys108, Lys109 and Arg123 in the alpha-subunit coupled with Lys22, Lys102, Lys116 and Arg117 in the beta-subunit, appears to be the primary determinant of its platelet-agglutinating activity. Conceivably, this interesting venom factor may provide a useful tool to study platelet agglutination by binding to the GPIb-IX-V complex.

Amino Acid Sequence↗
PubMedDate not supplied

Thimerosal dependent agglutination, a newly described blood bank problem.

A number of ABO grouping, Rh typing, antibody screening, and antibody identification problems are associated with chemicals in blood bank reagents. We describe a newly discovered agglutination phenomenon due to a thimerosal (Merthiolate)-dependent agglutinin found in the serum of a normal blood donor. Thimerosal is used as a preservative in several low-ionic strength reagents. This agglutination phenomenon is detected only in test systems (low-ionic-strength, albumin, saline, ficin treated test cells) in which test cells are incubated in the presence of thimerosal. Agglutination does not occur in the absence of thimerosal. The thimerosal-dependent agglutinin behaves like an IgG IgG autoantibody. There is no evidence that the thimerosal-dependent agglutinin is responsible for increased red cell destruction.

Adult↗
PubMed1991

Antibodies to histo-blood group substances A and B: agglutination titers, Ig class, and IgG subclasses in healthy persons of different age categories.

Isotypes and IgG subclasses of ABO antibodies from sera of 235 healthy blood donors were determined by an enzyme-linked immunosorbent assay (ELISA). Synthetic A and B trisaccharide-bovine serum albumin glycoconjugates were used for coating and monoclonal antibodies for the detection of heavy chain isotypes. Hemagglutination titers were determined in addition. Blood donors were between 20 and 67 years old, and at least 10 sera per 10-year age category and ABO blood group were included in this study. Antibody concentrations were expressed as a percentage of an internal standard, and sera with subclass-restricted anti-A and/or anti-B (anti-A/B) responses were used to normalize the ELISA values of IgG subclasses. A good correlation between the sum of the four subclasses and the total anti-A/B IgG values (rs = 0.81 for anti-A and 0.84 for anti-B) was obtained. IgG1 and IgG2 were the most predominant subclasses, but were found in various proportions in different individuals. Donor-to-donor variation exceeded age-related changes for all measured parameters. The correlation of anti-A IgM, IgG, IgA, and their sum with the agglutination titers was significant and revealed rs values of 0.70, 0.65, 0.65, and 0.80, respectively. For anti-B as well, the correlation of ELISA values with the agglutination titer was best when all three isotypes were added. We conclude that anti-A/B IgA, together with IgM and IgG, substantially contributes to the agglutination reaction. Potentially autoreactive antibodies were detected in sera of blood groups A, B, and AB.

ABO Blood-Group System↗
PubMedDate not supplied

Development of a batch-processed, passive latex agglutination test for cytomegalovirus.

BACKGROUND: The passive latex agglutination test is commonly used for the identification of cytomegalovirus IgG and IgM antibodies. This test is used because of its sensitivity, specificity, speed, and ease of performance, but it is unsuitable for large numbers of samples or for batch processing. STUDY DESIGN AND METHODS: To solve this problem, comparative studies to assess the cytomegalovirus passive latex agglutination test on microtiter plates were done with a new photometric particle agglutination method (PPAM) and an enzyme-linked immunosorbent assay as a control. RESULTS: A total of 3430 sera were tested using both the PPAM and enzyme-linked immunosorbent assay. A high degree (97.6%) of correspondence between the results of the tests was observed. The new PPAM was easier and faster to use (96 wells in 25 min). CONCLUSION: These results, as well as the possibility of adapting this method to a fully automated system, suggest that the PPAM could be an important contribution to the field of infection serology.

Antibodies, Viral↗
PubMed1999

Comparison of the performance of four microtube column agglutination systems in the detection of red cell alloantibodies.

BACKGROUND: The purpose of this study was to compare the performance of four currently available microtube column agglutination systems in the detection of red cell alloantibodies to that of the standard tube low-ionic-strength solution (LISS) indirect antiglobulin test (IAT) (tube LISS-IAT). STUDY DESIGN AND METHODS: In a comparative study, 172 sera, previously demonstrated to contain red cell alloantibodies, were tested in parallel by the tube LISS-IAT and three microtube column agglutination techniques (DiaMed-ID, Ortho BioVue, and Sanofi-Pasteur Scangel) and one affinity-adherence test system (Gamma ReACT). Tests were performed simultaneously by a single person on freshly thawed sera that had been frozen at -20 degrees C. RESULTS: The rate of detection of clinically significant alloantibodies (n = 154) in microtube column systems was very similar. One hundred forty-one sera (91.6%) reacted in the DiaMed-ID, 139 (90.3%) in the ReACT, 139 (90.3%) in the BioVue, and 142 (92.2%) in the Scangel. Only 117 (76.0%) of these sera reacted in the tube LISS-IAT. The detection rates for 18 antibodies of minor clinical significance (anti-M, -N, -P1, -Le(a), and -Le(b)) varied among the test systems: DiaMed-ID, 5 (28%); ReACT, 7 (39%); BioVue, 14 (78%); Scangel, 10 (56%); and tube LISS-IAT, 6 (33%). Antibody reactivity as determined by titer and score was very similar in all microtube column systems and higher in these systems than in the tube LISS-IAT. CONCLUSION: The sensitivity of all four microtube column systems in the detection of clinically significant red cell alloantibodies was similar and was markedly superior to that of the tube LISS-IAT. An individual cost-benefit analysis should be performed in every institution to decide whether a microtube column system should be applied. If so, the antibody screen in the microtube column agglutination system should ideally be performed in advance of the crossmatch to provide time to screen for compatible units.

Erythrocytes↗
PubMedDate not supplied

[Detection of Cryptosporidium oocysts in goat kid faeces: comparison of a latex agglutination test with three other conventional techniques].

Detection of Cryptosporidium oocysts from goat kid faeces: comparison of a latex agglutination test with three other conventional techniques. A quantitative latex agglutination test (QLAT) with monoclonal antibodies for the detection of Cryptosporidium oocysts in faeces was compared with 3 other conventional techniques: Heine staining on faecal smears (HS) giving semi-quantitative results (scores from 1 to 5), sucrose flotation on diluted faeces (SF) with results expressed in oocysts/g of faeces (opg), direct ELISA (DE) giving qualitative results. Goat kid unconcentrated faecal samples (234) from 8 farms were processed according to the 4 techniques. Data were analyzed with Win Episcope 1.0 and Testview 1.1 softwares. The oocyst outputs ranged from 100 000 (detection limit for SF) to 200 millions opg (mean: 15.2 millions opg). A very good agreement was recorded between QLAT and HS, SF, DE: Kappa values ranged between 0.82 and 0.90. When considering the samples exhibiting oocysts (or not) as positive (or negative) using both HS and SF (n = 219), the sensitivity and specificity of QLAT were respectively 95.1 and 96.0%. The lack of sensitivity was observed in faeces harboring a few oocysts (< or = 200 000 opg, scores < or = 2) whereas the lack of specificity was only observed in 3 samples originating from the same farm. A significant correlation was calculated between the percentage of agglutination in QLAT and the number of oocysts in SF or scores in HS (Spearman correlation ranging from 0.45 to 0.48, p < 0.001). QLAT is a rapid, simple and reliable tool for routine detection of Cryptosporidium oocysts in faeces.

Animals↗
PubMed1975

[Persistence of latex-agglutination inhibiting and neutralizing antibodies against enterovirus in the healthy population (author's transl)].

Sera of 122 persons were examined by the neutralization and latex-agglutination inhibition tests for antibodies against Polio type I, Coxsackie virus type A 9 and B 4, as well as Echo virus type 9. With increasing age there was a titre difference between latex-agglutination inhibiting and neutralizing antibodies, due to the earlier fall in the former. These antibodies, contrary to neutralizing ones, were in the lower titre region. The results confirmed the usefulness of the latex-agglutination inhibition test in the diagnosis of acute enterovirus infections.

Acute Disease↗
PubMed1980

Cell-cell recognition in yeast: purification of Hansenula wingei 21-cell sexual agglutination factor and comparison of the factors from three genera.

Trypsin digestion of Hansenula wingei 21-cells releases a protein (21-factor-T) that inhibits the agglutination of 21-cells by purified 5-agglutinin obtained from 5-cells by subtilisin digestion [Crandall, M. A. & Brock, T. D. (1968) Bacteriol. Rev. 32, 139-163]. We have purified this inhibitor 415-fold by ion-exchange chromatography, affinity adsorption to 5-cells, and gel permeation chromatography. The material shows a diffuse band, on polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate, with an apparent M(r) of 27,000. It has a pI of 3.8, is rich in acidic amino acids, contains 5% mannose and a trace of glucosamine, and is stable to reducing agents but is inactivated by heat. Zymolyase (beta1-->3-glucanase) digestion of 21-cells releases a similar inhibitor that, after purification, has a larger size than 21-factor-T. This 21-factor-Z appears to contain an additional portion that may serve to anchor 21-factor in the cell wall. Haploid cells of the yeasts Pichia amethionina and Saccharomyces kluyveri also show a constitutive sexual agglutination, and little or no crossreactivity is observed in heterologous mixtures. The agglutination factors in all three genera, however, have parallel properties; one cell type of each pair is heat stable and is inactivated by reducing agents (H. wingei 5-cells, P. amethionina alpha-cells, and S. kluyveri 16-cells), and the other is heat labile and is unaffected by reducing agents H. wingei 21-cells, P. amethionina a-cells, and S. kluyveri 17-cells). Because S. kluyveri 16-cells respond to Saccharomyces cerevisiae alpha-factor with the typical morphogenetic change of a mating half-reaction, the heat-stable agglutinin appears related to the S. cerevisiae a mating type and the heat-labile factor to the S. cerevisiae alpha mating type.

Ascomycota↗
PubMed2000

Detection of anti-GM1 ganglioside antibodies in patients with neuropathy by a novel latex agglutination assay.

Highly elevated titers of serum anti-GM1 ganglioside antibodies are closely associated with multifocal motor neuropathy, but low titers are commonly present in normal individuals or other diseases. Current systems for measuring anti-GM1 antibodies utilize the enzyme-linked immunosorbent assay (ELISA), in which serum dilutions are tested for binding to excess antigen immobilized on the surface of microwells. The ELISA system, however, is relatively time consuming, labor intensive, and costly, in addition to being prone to methodological variability. We have developed a novel agglutination assay for the detection of anti-GM1 antibodies, utilizing GM1 ganglioside-coated latex beads. In contrast to the ELISA system, antibody titers may be quantified by testing for agglutination using latex beads coated with decreasing amounts of antigen. The agglutination assay compares favorably to the ELISA system in sensitivity and specificity, but is considerably less costly and takes only a few minutes to perform.

Amyotrophic Lateral Sclerosis↗
PubMed2002

A comparison of spiral computed tomography and latex agglutination D-dimer assay in acute pulmonary embolism using pulmonary arteriography as gold standard.

OBJECTIVE: To compare the diagnostic accuracy of contrast medium enhanced spiral computed tomography of the pulmonary arteries (s-CTPA) and a latex agglutination D-dimer assay in patients with suspected acute pulmonary embolism (PE) by using pulmonary arteriography (PA) and clinical follow-up as reference method. DESIGN: Ninety hemodynamically stable patients with symptoms of acute pulmonary embolism were prospectively evaluated with s-CTPA and pulmonary arteriography (PA) within 24 h from admission. Plasma D-dimer levels on admittance were analyzed using a rapid latex agglutination D-dimer assay. The outcome of D-dimer concentrations in plasma below 0.25 and 0.5 mg/l was studied. RESULTS: All PA and s-CTPA investigations were regarded as of acceptable diagnostic quality in a consensus reading. Thirty-three patients had a positive PA (37%). Three patients had false negative and two patients had false positive s-CTPA findings. s-CTPA had 91% sensitivity, 96% specificity, 94% positive predictive value (PPV) and 95% negative predictive value (NPV). The sensitivity and specificity for D-dimer below 0.5 mg/l were 79 and 88%, respectively. The PPV and NPV were 81 and 87%. If a cut-off level of 0.25 mg/l was used the corresponding figures were 91, 65, 63 and 92%. CONCLUSION: s-CTPA has a higher sensitivity and specificity than latex agglutination D-dimer. A cut-off level of 0.25 mg/l can be used as screening method, but s-CTPA must be performed to exclude false positive cases.

Adolescent↗