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PubMed1992

Botrocetin agglutination of rat megakaryocytes: a rapid method for megakaryocyte isolation.

Botrocetin isolated from the venom of Bothrops jaracara has been shown by others to induce binding of von Willebrand Factor to glycoprotein Ib and thereby produce platelet agglutination in a wide range of animal species. We have found that botrocetin also facilitates the agglutination of megakaryocytes and have used this property to develop a method to isolate megakaryocytes from rat bone marrow. When botrocetin is added to a mixture of rat bone marrow, rat platelets, and rat plasma, coagglutination of megakaryocytes and platelets occurs. The agglutinated complexes, containing > 95% of the megakaryocytes, may then be separated from the remaining marrow cells by filtration. Megakaryocytes account for 39% (range 30%-48%) of the isolated cells and 83% (range 77%-88%) of the isolated cell mass. This method allows the virtually complete removal of megakaryocytes from bone marrow as well as their isolation to a high degree of purity. It should provide a useful, inexpensive, general method for the rapid isolation of megakaryocytes from multiple, small marrow samples from a wide range of species.

Animals↗
PubMed2006

H-deficient Bombay and para-Bombay red blood cells are most strongly agglutinated by the galactophilic lectins of Aplysia and Pseudomonas aeruginosa that detect I and P1 antigens.

The galactophilic lectins Aplysia gonad lectin (AGL) and Pseudomonas aeruginosa lectin (PA-IL), which detect human I and P1 RBC antigens, were examined for hemagglutination of H+ (group O and B) and H-deficient (Bombay and para-Bombay phenotype) RBCs. The results were compared with those obtained using two other galactophilic lectins, Maclura pomifera lectin (MPL) and Arachis hypogaea (peanut) agglutinin (PNA), which share T-antigen affinity, and two fucose-binding H-specific lectins, Ulex europaeus (UEA-I) and Pseudomonas aeruginosa lectin (PA-IIL), as well as with those achieved with anti-I serum. The results revealed that, in contrast to UEA-I and PA-IIL, which preferentially agglutinated H+ RBCs, and to MPL and PNA, which similarly agglutinated all examined RBCs, AGL, PA-IL, and the anti-I serum agglutinated the H-deficient RBCs more strongly than did the H+ RBCs. These findings could be attributed to increased levels of I and P1 antigens on those RBCs resulting from the use of the free common H-type 2 precursor for their synthesis. Since both PA-IL and PA-IIL are regarded as potential pathogen adhesins, it would be interesting to statistically compare the sensitivities of individuals of H+ and H-deficient RBC populations to P. aeruginosa infections.

ABO Blood-Group System↗
PubMed1991

Comparison of latex agglutination and polyacrylamide gel electrophoresis with enzyme linked immunosorbent assay for detecting human rotavirus in stool specimens.

One hundred and forty five stool samples from children below 2 years of age, hospitalized with diarrhea were tested for rotavirus antigen by enzyme linked immunosorbent assay (ELISA), latex agglutination test using commercially available kit Rotastat (Ranbaxy Diagnostic, India) and by polyacrylamide gel electrophoresis. Twenty eight samples were positive for the virus antigen by all the three assay systems. The sensitivity of latex agglutination (LA) and polyacrylamide gel electrophoresis (PAGE) was 91.4% (32/35) and 80% (28/35), respectively; the corresponding specificity was 98.18% (108/110) and 100% (110/110), respectively. Latex agglutination was the least complex, required the least amount of apparatus and provided a result within a short time. It showed a high specificity and a reasonable amount of sensitivity and the results correlated well with ELISA and PAGE.

Antigens, Viral↗
PubMed2006

[Development of the simple method for measurement the content of Cry j 1 in the air by latex agglutination test].

BACKGROUND: In order to develop an easy method for measure the airborne C. japonica pollen allergen (Cry j 1) without any utensils or a special apparatus, we examined latex agglutination test for the purpose. METHODS: The latex beads were reacted with the anti-Cry j 1 polyclonal antibody, and the beads were blocked with bovine serum albumin (sensitized beads). The measurement is as follows; two-fold diluted series of solutions of airborne samples are reacted with the sensitized beads, and the results are examined after 2 hrs. RESULTS: Agglutination was recognized with 5ng/ml Cry j 1 standard solution and 20pg/m3 of airborne samples. CONCLUSION: We have developed a simple method for measure the airborne Cry j 1 by use of latex agglutination test. The strong point of the method is 1) measurement is easy and does not affect the numbers of pollen, 2) do not need any utensils or a special apparatus, 3) measure the disease causative materials themselves.

Air Pollutants↗
PubMed1990

[Study of the latex agglutination reaction using a light-scattering method].

Light scattering techniques as a method for monitoring latex agglutination test were under study. The model system, employed in the studies, consisted of a suspension containing polystyrene latex microspheres with tetanus antitoxin immobilized on them (carrier particles) and antitetanus serum (3000 IU). The studies helped define approaches to the choice of the optimal conditions for monitoring the latex agglutination test. Light scattering method has proved to be highly sensitive, its sensitivity essentially surpassing that of slide tests and permitting the detection of early agglutination stages.

Latex Fixation Tests↗
PubMed1975

[Formation and persistence of latex-agglutination-inhibiting IgG- and IgM-antibodies in rabbits inoculated with poliomyelitis, Coxsackie and encephalomyocarditis viruses (author's transl)].

The formation and presistence of virus specific IgG- and IgM-antibodies produced in rabbits after a single dose of poliomyelitisvirus, coxsackievirus or encephalomyocarditisvirus were tested comparatively by latex-agglutination-inhibition test, hemagglutination-inhibition test and neutralization test. The animals were immunized either intravenously alone or with Freund's complete adjuvant. The IgM-antibodies were only detectable for a short time by the three tests. In the IgG-fraction and in the unfractionated serum the neutralizing antibodies persisted without any loss in titer during the period observed, whereas the latex-agglutination-inhibiting and hemagglutination-inhibiting antibodies showed a significant decrease. The hemagglutination-inhibiting antibodies required 23 weeks (Figure 5), the latex-agglutination-inhibiting antibodies 7 weeks (without Freund's complete adjuvant; Figure 1 and 3) or about 16 weeks (with Freund's complete adjuvant; Figure 2, 4 and 5) respectively to decline to 25% of their maximum titer.

Animals↗
PubMed1991

The lack of utility of the latex agglutination test for detection of Candida antigen in the diagnosis of systemic candidiasis.

The Ramco latex agglutination test in the diagnosis of systemic candidiasis was utilized for 11 serum samples from 10 patients with systemic candidiasis, 21 serum samples from patients colonized with Candida species, and 20 control serum samples from patients with stable medical problems and no evidence of Candida albicans infection. This study was double-blind and the results of the latex agglutination test did not influence the decision for antifungal therapy. Nine of 10 patients with systemic candidiasis had positive titers (greater than or equal to 1:4); however, these were determined only 1 to 5 days before culture positivity. Nine of 21 (43%) of colonized patients were falsely positive (greater than or equal to 1:4) and all of the control samples were negative. The Ramco latex agglutination test was unreliable and inconsistent in this small sample group to establish an early diagnosis of systemic candidiasis.

Antigens, Fungal↗
PubMed1991

[The use of the latex agglutination reaction for the diagnosis of a Brucella infection].

The authors have developed the optimum conditions for the preparation of antigenic diagnosticum based on latex manufactured in the USSR. To sensitize latex with the diameter of microspheres equal to 0.83 microns, Brucella polysaccharide was used in a dose of 100 micrograms/ml. As stabilizer, polyvinylpyrrolidone at a concentration of 0.1% was used. The specificity and sensitivity of the diagnosticum were studied in analysis of serum samples taken from 102 healthy donors and patients with infectious diseases of nonbrucellar etiology and from 120 patients with different forms of brucellosis. The specificity of the diagnosticum was found to be 94.1% and its sensitivity, 77.5%. Comparative study of the latex agglutination test with other serological tests showed that the former test is highly effective both in acute and chronic forms of the disease. A high degree of correlation between the agglutination test, Coombs' test, the passive hemagglutination test and the latex agglutination test was established (r = 0.83, 0.72 and 0.62, respectively).

Antibodies, Bacterial↗
PubMed1991

Dimeric ristocetin flocculates proteins, binds to platelets, and mediates von Willebrand factor-dependent agglutination of platelets.

Ristocetin in aqueous solution dimerizes with an equilibrium dissociation constant of 5.0 x 10(-4) M, i.e. approximately 1.1 mg ml-1 (Waltho, J.P., and Williams, D. H. (1989) J. Am. Chem. Soc. 111, 2475-2480). At concentrations of about 1.0 mg ml-1 ristocetin flocculates many proteins, lyses platelets and, in the presence of von Willebrand factor, agglutinates both fresh and formalin-fixed platelets. Because ristocetin exists as both monomeric and dimeric species, we sought to determine which of these forms flocculates proteins and agglutinates platelets. We found that: 1) the initial rate of flocculation of certain proteins, 2) the initial rate of agglutination of formalin-fixed platelets, and 3) the binding of ristocetin to formalin-fixed platelets are higher order solely with respect to the concentration of ristocetin dimers. As to the operative mechanism, it appears that bifunctional dimers cross-link proteins that possess multiple copies of a common recognition site. Preliminary evidence indicates that a recognition site is a beta-turn of the form X-P-G-X'.

Afibrinogenemia↗
PubMedDate not supplied

A comparison of two macroscopic platelet agglutination assays for von Willebrand factor.

Plasma drawn from 22 patients with a broad range of von Willebrand factor (vWF) levels, including one patient with rare, severe type III von Willebrand's disease, was used to investigate the adaptability of the Bio/Data Corp. vW Factor Assay, a ristocetin cofactor-based kit developed for use with a platelet aggregometer, to a macroscopic platelet agglutination technique. The results were compared to those obtained using the General Diagnostics von Willebrand Factor Assay, another ristocetin cofactor-based kit developed for use as a macroscopic platelet agglutination procedure. No significant difference (p greater than 0.5) existed between results obtained by these two kits. The mean and standard deviation of the vWF levels (as percent of normal) were 132 +/- 66.7 by the General Diagnostics kit and 133 +/- 59.0 by the Bio/Data Corp. kit. The correlation coefficient (r) was 0.81. The Bio/Data Corp. kit is more costly per test than the General Diagnostics kit. However, it was shown to be compatible with the macroscopic agglutination technique and required no modification to be used in that manner.

Female↗
PubMed1985

Immunochemical qualitative latex agglutination test for pancreatic lipase in serum evaluated for use in diagnosis of acute pancreatitis.

In 417 patients (213 men, 204 women) consecutively hospitalized with acute abdominal pain we evaluated the clinical usefulness of a latex-agglutination test at admission to screen for concentrations of pancreatic lipase (EC 3.1.1.3) in serum greater than 300 micrograms/L. The diagnoses of acute pancreatitis (in 25 patients, 6%) and other diseases were made without knowledge of the results of the latex test or of quantification of pancreatic lipase in the serum by enzyme immunoassay. In the latex assay, when agglutination was taken as a positive test for acute pancreatitis, we found a diagnostic efficiency of 0.986 (95% confidence limits: 0.971-0.997) for acute pancreatitis. The predictive value of a positive latex test result with respect to acute pancreatitis was 0.807 (0.625-0.926); the predictive value of a negative test was 1.000 (0.991-1.000). Six patients had false-positive test results. No false-negative test results were found by enzyme immunoassay. We conclude that the latex agglutination test is useful as an emergency test for diagnosis of acute pancreatitis in patients with acute abdominal pain; negative results virtually exclude acute pancreatitis.

Acute Disease↗
PubMed1987

Three latex agglutination tests compared with gram staining for the detection of bacteria in cerebrospinal fluid.

Three commercially available latex agglutination tests and the Gram stain were evaluated for their ability to detect Haemophilus influenzae, Streptococcus pneumoniae and Neisseria meningitidis in cerebrospinal fluid (CSF) specimens from 160 patients suspected of having meningitis. Bactigen detected 86% of all culture-positive CSF specimens, whereas Wellcogen detected 80%, Bio Merieux 59% and Gram stain 94%. Bactigen and Wellcogen detected 84% of cases of meningitis caused by H. influenzae, whereas Bio Merieux detected 53% and Gram stain 90%. Of the 27 cases of Strept. pneumoniae meningitis, all were detected by Gram stain, 96% by Bactigen, 89% by Wellcogen and 74% by Bio Merieux. For N. meningitidis, which included four isolates of group B and one group W135, detection was 80% with the Gram stain, 40% with Bactigen and 20% with Wellcogen. False-positive results were observed with Bactigen and Bio Merieux respectively on 5% of culture-positive CSF specimens for organisms other than the test organisms. With the exception of faulty batches of individual kits of Bactigen (H. influenzae B) and Wellcogen (Strept. pneumoniae) respectively, false-positives were not obtained on the 63 culture-negative CSF samples. Bactigen and Wellcogen were equally efficacious in detecting H. influenzae B. Bactigen was marginally superior to Wellcogen for the detection of Strept. pneumoniae. Bio Merieux was the least sensitive of the three latex agglutination kits tested. The Gram stain was found to be at least as sensitive or superior to latex agglutination for detecting H. influenzae B, Strept. pneumoniae and N. meningitidis in CSF specimens from patients with bacteriologically proven meningitis.

Cerebrospinal Fluid↗
PubMed1989

[A modified method of latex agglutination in capillaries for the detection of meningococcal antigens].

The suggested microvariant of the latex agglutination test in glass capillaries is not inferior to the traditional latex agglutination on the glass in sensitivity and specificity. The microvariant consists in the following: glass capillaries 0.8 mm in diameter 10 cm long are filled with the pathological material (3/5 of the tube length) and with the diagnostic latex preparation (1/5 of the tube length) so that no air hole is formed between them. The time of the reaction may be prolonged to 20-30 min without deteriorating its specificity. The results are recorded using a 4+ system, similarly as with the latex agglutination on the glass. The described modification appears to be useful for practical laboratories: it saves the latex preparation 4-fold and takes 2 times less pathologic material.

Antigens, Bacterial↗
PubMed1989

Relationship of human macrophage agglutination factor to other fibronectins.

Human macrophage agglutination factor (MAggF) is a lymphokine with a number of biochemical and immunochemical similarities to the fibronectins (FN). The present study was undertaken to define this relationship more exactly. Peripheral blood mononuclear cells (PBMC) and cloned human CD4+T-cell lines synthesized both MAggF and immunoreactive FN de novo following antigen or mitogen activation. MAggF produced under those conditions co-purified with an immunoreactive FN on gelatin-affinity and gel-filtration chromatography. This FN had a consistently slightly smaller relative molecular mass than plasma FN on both immunoblotting and gel filtration. Purified human MAggF was extremely active: it agglutinated human monocytes at fM concentrations and was up to 2,000,000 times more active in agglutinating mononuclear phagocytes than other purified FN. The action of MAggF on monocytes was dependent on at least two antigenically distinct cell-surface FN receptors. We suggest that the extreme activity of MAggF is a result of co-operative interactions between FN domains on the MAggF molecules and multiple distinct classes of FN receptors on responding cells.

Animals↗
PubMed1988

Detection of rinderpest antigen by latex agglutination test.

A slightly modified latex agglutination test was applied for detection of rinderpest antigen. The antigen was added to sensitized latex particles in the presence of hyperimmune antiserum to facilitate agglutination. Out of 129 samples tested by latex agglutination (LA), solid phase aggregation of coated erythrocytes (SPACE), reverse phase passive haemagglutination (RPHA) and counter immunoelectrophoresis (CIE) test, 86.0, 86.8, 84.4 and 79.8 per cent, respectively, were found positive.

Antigens, Viral↗
PubMed1985

Agglutination of platelet-type von Willebrand's disease platelets by bovine von Willebrand factor.

It has been shown that platelets from patients with platelet-type von Willebrand's disease (vWD) agglutinate upon the addition of human von Willebrand factor (vWF) in the absence of ristocetin or botrocetin, suggesting that platelet membrane receptors for human vWF is abnormal. The present work reports the platelet agglutinability on stimulation with bovine vWF in platelet-type vWD. Platelets in patient platelet-rich plasma or washed platelet suspensions and patient platelets treated with formalin agglutinated in the presence of markedly lower concentrations of bovine vWF than those required for normal platelets. This finding provides additional evidence that platelet-type vWD platelets have abnormal expression of binding sites for vWF on their surface, and supports that platelet receptors for bovine vWF are identical or very close to those for human vWF.

Animals↗
PubMed1977

Inhibition of ristocetin-induced platelet agglutination by vancomycin.

Ristocetin and vancomycin are structurally similar glycopeptide antibiotics. Both vancomycin and ristocetin in high concentrations (3.0 mg/ml) cause the precipitation of fibrinogen, plasminogen, and IgG from platelet-poor plasma (PPP). In contrast to ristocetin, vanomycin (0.5-1.5 mg/ml) does not agglutinate platelets in normal platelet-rich plasma (PRP) or formalin-treated platelets in the presence of normal PPP. Preincubation of vancomycin (0.5-1.25 mg/ml) with normal PRP, von Willebrand platelets in normal PPP, or formalinized platelets results in inhibition of platelet agglutination induced by ristocetin (0.7-1.25 mg/ml) or ristocetin and normal PPP. This inhibition can be overcome by increasing the final concentration of ristocetin in the platelet suspension. Preincubation of formalin-treated platelets with the major fraction obtained by carboxymethyl-Sephadex C-50 chromatography of commercial vancomycin also results in inhibition of agglutination induced by ristocetin and normal PPP. Incubation with vancomycin (1.25 mg/ml) does not interfere with von Willebrand factor (vWF) or factor VIII coagulant activities in normal PPP or in Sepharose 4B void volume fractions of PPP. These results indicate that vancomycin interacts with normal, von Willebrand, and formalin-treated platelets and inhibits the binding of ristocetin (or ristocetin-vWF complexes).

Binding, Competitive↗
PubMed1986

Agglutination of intravenous lipid emulsion ('Intralipid') and plasma lipoproteins by C-reactive protein.

Isolated human C-reactive protein (CRP), or CRP in acute phase serum, produced in-vitro agglutination ('creaming') of the intravenously administered lipid suspension 'Intralipid'. CRP also produced similar agglutination of isolated normal very low density lipoproteins (VLDL). Agglutination in both cases was calcium-dependent and inhibitable by phosphoryl choline. These findings have important implications for patients receiving intravenous lipid suspensions and may be relevant to the pathogenesis of the fat embolism syndrome following trauma.

Acute-Phase Reaction↗