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Detection of rotavirus in faeces by latex agglutination.

Human rotavirus (HRV) in faeces of patients may be readily detected with high sensitivity and specificity using latex agglutination (LA) on a glass slide by making use of the cross-reactivity of anti-calf rotavirus (CRV) antibody. Latex particles were coated with anti-CRV immunoglobin. The antibody coated particles (AC-L) are specifically agglutinated by both CRV and HRV, and the agglutination is evident macroscopically within a minute. To examine the sensitivity and reliability of the LA method compared to other methods, HRV in faecal extracts of 48 infants with acute gastroenteritis was sought by the LA, reversed passive haemagglutination (RPHA) and electron microscope (EM) methods. Samples positive by the EM method were all positive by the LA method, and samples negative by EM were all negative by LA. The LA method is suitable for application as a simple clinical diagnostic test.

Child, Preschool↗

A critical analysis of commercially available latex particle reagents for C-reactive protein (CRP) slide agglutination tests.

C-reactive protein (CRP) was assessed in pediatric serum samples using different commercial latex reagents, which were analyzed for species origin of the coating antibodies, homogeneity and density of the latex particles, and prozone agglutinating capacity. All reagents correctly agglutinated the positive and negative control sera. The antibodies coating the particles differed with regard to species origin: one was coated with rabbit, one with horse and goat, one with horse, goat, rabbit and swine, while the reference reagent had horse, goat and rabbit antibodies. Only the monospecies specific antibody-coated latex showed obvious prozoning; this reagent also had the smallest and most homogenous latex particles and showed the most clear-cut reactions. False agglutination was observed at 7-26% according to quantitation with the spot immunoprecipitate assay, which compared favorably with radial immunodiffusion measurements. The lowest percentage of false readings was noted for the rabbit antibody-coated particles; the highest for the reagent with particles coated using antibodies from 4 different species. No reagent had satisfactory precision for the low positive sera between 10 and 40 mg CRP/l.

C-Reactive Protein↗

Detection of adenovirus type41 in stool samples by a latex agglutination method.

We have developed a simple agglutination (LA) method for the detection of enteric adenovirus (EAd) in stool samples from infants with acute gastroenteritis. Ad type 41 (Ad41) was detected with high sensitivity and specificity by a slide agglutination test using latex particles coated with antiAd41 antibody (LA-antiAd41). The agglutination of LA-antiAd41 with Ad41 on a glass slide was evident macroscopically within 2 min. The sensitivity of the LA method was four times higher than that of the EM method.

Adenoviruses, Human↗

Simplified conjugation chemistry for coupling peptides to F(ab') fragments: autologous red cell agglutination assay for HIV-1 antibodies.

The rapid whole blood test, developed for the detection of circulating antibodies to human immunodeficiency virus type 1 (HIV-1), is based on agglutination of autologous red blood cells using an anti-human glycophorin antibody conjugated to the HIV-1 immunodominant epitope of gp41 (579-613). A simplified procedure for preparing antibody-peptide conjugates for use in the autologous red cell agglutination test is described. F(ab')2 fragments of the anti-glycophorin antibody were prepared by pepsin digestion and reduced to F(ab') fragments with the use of tri-n-butylphosphine (TBP). This permitted the simultaneous reduction of the F(ab') fragments and coupling of a bromoacetyl derivative of the synthetic immunodominant peptide gp41 (579-613) [Cys-Acm 598, Lys-BrAc 604] containing epsilon-bromoacetyl-lysine at residue 604 to the resultant F(ab') fragment. Conjugation to the F(ab') fragment resulted in a stable thio-ether linkage between the peptide Lys-604 and the inter heavy chain cysteines of the F(ab'). The resultant F(ab')-peptide conjugate was comparable to the previously described disulfide coupled conjugate when used in the autologous red cell agglutination test. This simplified conjugation chemistry may also be useful for the development of reagents for FACS analysis as well as targetted vaccines.

Amino Acid Sequence↗

A kinetic model of the agglutination process.

We propose a kinetic model of the aggregation process in a system consisting of two different types of particles. Aggregating particles (cells) are polyvalent and bear on the surface a huge number of binding sites for the other type of particles, ligands. The ligand is bivalent and has two identical active sites for binding to cells. The cross-linking of the cells by the ligands causes the aggregation phenomenon called agglutination. We obtained the analytical solution of this model task describing the time dependence of the aggregate mean size versus the composition of the system. The comparison of the analytical solution with the experimental data for the agglutination of bacterial cells by bivalent antibodies shows that the main factors affecting agglutination were correctly taken into account.

Bacterial Adhesion↗

Evaluation of a commercial latex agglutination test for detecting antibodies to cytomegalovirus in organ donors and transplant recipients.

A commercial latex agglutination test was evaluated for use in detecting CMV antibody in organ donors and transplant recipients. When compared with indirect and competitive enzyme-linked immunosorbent assays (ELISAs) and complement fixation tests (CFT), the test gave concordant results with 96.6% of sera. The latex agglutination test was more sensitive than CFT but less sensitive than ELISA. The major advantage of the latex agglutination test was its simplicity and rapidity. Taking only 8 min to perform, it is very suitable for testing sera from organ donors, since the time available for such tests is short because of the finite ischaemic time, particularly for liver and heart transplantation.

Antibodies, Viral↗

A latex agglutination test for the detection of canine parvovirus and corresponding antibodies.

Latex particles coated with rat monoclonal antibodies directed against a canine parvovirus are agglutinated by the virus antigens. In this study, the reaction was quantitated by a technique which had been described as immunoassay by particles counting. The method is as sensitive as a radioimmunoassay with the same antibody, but has the advantages of simplicity and safety. It allows the detection of parvoviral antigens to 4 ng/ml. Incubation of these antigens with specific antibodies resulted in inhibition of the agglutination reaction. By this procedure the presence of antibodies against canine parvovirus can be detected to a concentration of 150 ng/ml. The system can be easily automated, thus increasing reproducibility. This latex agglutination technique can be performed as a slide test; and although this method is 4-5 times less sensitive, it could be useful in field studies, for the detection of the viral antigens and the corresponding antibodies.

Animals↗

Agglutination of human O erythrocytes by influenza A(H1N1) viruses freshly isolated from patients.

The hemagglutinin titers of 10 influenza A (H1N1) viruses were examined using the erythrocytes of several species. Human O erythrocytes showed the highest agglutination titer to the viruses, whereas chicken erythrocytes showed a low titer. These findings were noted for at least 10 passages by serial dilutions of the viruses in Madin-Darby canine kidney (MDCK) cells. All influenza A(H1N1) viruses, plaque-cloned directly from throat-washing specimens of patients, also agglutinated human O but not chicken erythrocytes. The results of a hemadsorption test indicated that chicken erythrocytes possess less affinity to MDCK cells infected with the A/Osaka City/2/88(H1N1) stain than to those infected with the A/Yamagata/120/86(H1N1) strain which is used as an inactivated influenza vaccine in Japan. However, there were no significant differences between the A/Osaka City/2/88 and the A/Yamagata/120/86 strains in the hemagglutination inhibition test. Since human O erythrocytes have high agglutination activity to influenza A(H1N1) and also to A(H3N2) and B viruses in MDCK cells, these erythrocytes may be useful for the serological diagnosis of influenza.

Adult↗

Comparison of counterimmunoelectrophoresis and latex particle agglutination in the detection of Streptococcus pneumoniae in blood cultures.

Blood cultures from 140 patients with chest infections were examined for the presence of pneumococcal antigen by both latex particle agglutination and counterimmunoelectrophoresis. Twenty-three samples (15 patients) were positive for pneumococcal antigen by counterimmunoelectrophoresis, of which 21 samples (13 patients) yielded Streptococcus pneumoniae in culture. Thirty-three samples (20 patients) were positive by latex particle agglutination, of which 24 samples (15 patients) were culture positive. The sensitivities and specificities were calculated and it was concluded that latex particle agglutination was the better test for the determination of the presence of pneumococcal capsular polysaccharide.

Antigens, Bacterial↗

Effects of ultrasound on agglutination and aggregation of human erythrocytes in vitro.

A new experimental approach has shown that human erythrocytes of different blood groups were induced to form more agglutinates at a sound pressure of 70-240 kPa in vitro than the control erythrocytes. Similar effects were observed for alcian blue and dextran stimulated aggregation and for spontaneous aggregation. The increase of agglutination or aggregation was reversible. Heating and acoustic cavitation were shown not to be responsible for this effect. Bulk fluid movement produced by ultrasound irradiation appeared to cause the described phenomenon. Possible underlying mechanisms connecting the acoustic streaming and agglutination or aggregation behaviour of the cells are proposed.

Alcian Blue↗

Agglutination activity of Limulus polyphemus coagulogen following limited proteolysis.

1. A 14 kDa protein with cell agglutination properties has been purified from endotoxin-activated L. polyphemus amebocyte lysate. Amino terminal sequence analysis indicates that this protein corresponds to a proteolytically cleaved product (coagulin) of coagulogen. 2. Similar cell agglutination activity can be generated, in vitro, by proteolytic cleavage of the coagulogen with either trypsin, endogenous protease or an alpha 2M/enzyme complex isolated from amebocytes. 3. Studies with [125I]-labeled coagulogen showed that only coagulin, not the intact coagulogen, binds to rabbit erythrocytes and formalin-fixed amebocytes. 4. The cell agglutination activity of coagulin towards erythrocytes was not inhibited by various sugars tested, and was not Ca(2+)-dependent. 5. These findings suggest that coagulogen and coagulin are reminiscent of their mammalian counterparts, fibrinogen and fibrin, in their clotting and relative adhesive properties.

Amino Acid Sequence↗

Evaluation of latex agglutination and microtube coagulase tests for detection of Staphylococcus aureus.

In a blind study, a latex agglutination test (Serostat Staphylococcus, Scott Laboratories) and a microtube coagulase test (Staphase, API) were evaluated for their ability to detect Staphylococcus aureus. Of 289 isolates of catalase-positive, gram-positive cocci, 122 were identified as S. aureus based on positive reactions in at least three of the following tests: tube coagulase, slide coagulase, DNase production, or anaerobic fermentation of mannitol. The latex agglutination test gave positive reactions for all S. aureus isolates and 10 (6%) non-S. aureus isolates. The slide coagulase test was positive for 121 S. aureus isolates and three (2%) non-S. aureus isolates. The microtube coagulase test detected 53, 90, and 98% of the S. aureus isolates after 2, 4, and 24 hr, respectively. In contrast, the conventional tube coagulase test detected 97% of the S. aureus isolates after 2 hr, and 98% after 4 and 24 hr. Two isolates of S. aureus gave negative tube coagulase reactions at 37 degrees C, but positive reactions at room temperature after 24 hr. The combination of tube and slide coagulase tests provided the most reliable results. The slide and tube coagulase tests gave more reliable results than the latex agglutination and microtube coagulase tests, respectively.

Coagulase↗

Effects of female sex steroids on concanavalin A-mediated agglutination of hepatocytes from nonregenerating and regenerating rat liver and hepatic tumor marker enzymes.

Effect of treatment of female rats with an oral contraceptive agent (OCA), Ovulen-50, for 7 weeks on agglutination of hepatocytes with concanavalin A (con A) and activities of certain tumor marker enzymes were examined to find out if OCA treatment is related to preneoplastic or neoplastic processes. Hepatocytes from regenerating and nonregenerating livers of control female rats showed negligible agglutination with Con A, whereas hepatocytes from non regenerating but not from the regenerating livers of female rats treated with a combination of 5 micrograms ethinyl estradiol and 100 micrograms ethynodiol diacetate showed agglutination. Of the tumor marker enzymes such as hepatic glucose 6-phosphatase, gamma-glutamyl transpeptidase (gamma-GT), and arginase examined in the liver, only gamma-glutamyl transpeptidase showed a significant increase in activity in the steroid-treated rats. Plasma alkaline phosphatase activity was also higher in the treated animals. However, the magnitude of the changes observed was relatively small and perhaps unrelated to the neoplastic process.

Animals↗

Hydroquinone, a control agent of agglutination and adherence of Streptococcus mutans induced by sucrose.

Hydroquinone was found to alter agglutination of Streptococcus mutans induced by sucrose. The newly formed agglutination product produced by hydroquinone does not kill this cariogenic bacterium and the formation is reversible. The agglutination altering activity of hydroquinone seems to be specific for strains of S. mutans. As a result, hydroquinone inhibits sucrose-induced adherence of S. mutans.

Anti-Bacterial Agents↗

Latex agglutination for bacterial antigens and meningococcus PCR: two useful tools in legal sudden deaths.

Bacterial infections are considered to be a major cause of sudden deaths. The recognition of infections caused by Neisseria meningitidis is an essential duty of medicolegal offices due to the risk of secondary cases. Since other microorganisms, such as Haemophilus influenzae and Streptococcus pneumoniae, are also involved in infectious sudden deaths, the identification of the pathogen responsible for death is essential in order to establish a positive diagnosis while also preventing secondary meningococcal cases. However, because of the unreliability of culture methods used for autopsy specimens and the fragile nature of the microorganisms, other techniques were used. In this study, the detection of specific antigens of N. meningitidis (serogroups A, B, C, Y and W135), H. influenzae type b, S. pneumoniae and Group B Streptococcus was undertaken in 40 samples from sudden death cases in legal procedures with a latex agglutination test. In addition, a meningococcus polymerase chain reaction (PCR) assay (ctrA, crgA and siaD genes) was also used as a corroboration method for positive N. meningitidis agglutinations. Eleven cases of sudden death were confirmed to be due to meningococcus while one case was confirmed to have been caused by H. influenzae type b fulminant epiglottitis. Rapid laboratory diagnosis of meningococcal infection allowed contacts management and notification to the health authorities. From the point of view of the authors, forensic diagnosis of unascertained deaths should include latex agglutination and meningococcus PCR when a fulminant infection by N. meningitidis or H. influenzae is suspected as well as in deaths where the cause is unclear.

Adolescent↗

The C-terminal peptide of thrombospondin-1 stimulates distinct signaling pathways but induces an activation-independent agglutination of platelets and other cells.

A peptide from the C-terminal domain of thrombospondin-1 (4N1-1) has been proposed to stimulate platelet aggregation by a novel mechanism involving both an activation-independent agglutination and an activation-dependent, glycoprotein (GP) IIb/IIIa-mediated aggregation which involves GPVI signaling but does not involve CD47. The present study demonstrates that 4N1-1 stimulated a different pattern of signal transduction pathways than the GPVI agonist convulxin. Furthermore, 4N1-1-induced platelet aggregation was activation-independent and not dependent on GPVI or GPIIb/IIIa. Interestingly, 4N1-1 also stimulated activation-independent agglutination of different megakaryocytic and non-megakaryocytic cells. 4N1-1-induced cell agglutination but not platelet signaling was inhibited by anti-CD47 antibodies.

Antigens, CD↗

Treatment of vaginal agglutination associated with chronic graft-versus-host disease.

OBJECTIVE: To describe the presentation and clinical course of two patients after development of vaginal agglutination associated with chronic graft-versus-host disease. DESIGN: Case report. SETTING: Tertiary referral center for pelvic reconstructive surgery. PATIENT(S): Two patients with the diagnosis of chronic graft-versus-host disease who later developed vaginal agglutination requiring treatment. INTERVENTION(S): Surgical lysis of vaginal adhesions and postoperative use of vaginal dilators. MAIN OUTCOME MEASURE(S): Successful treatment of vaginal adhesions. RESULT(S): Both patients underwent successful surgical lysis of vaginal adhesions and maintained vaginal patency with postoperative use of vaginal dilators. CONCLUSION(S): Prompt diagnosis of vaginal agglutination in patients with chronic graft-versus-host disease and appropriate surgical correction of this complication rather than prophylaxis is the correct treatment course.

Chronic Disease↗

A novel ultrasound-enhanced latex agglutination test for the detection of antibodies against Mycobacterium tuberculosis in serum.

A novel ultrasound-enhanced latex agglutination test is described for the detection of serum antibodies against Mycobacterium tuberculosis. The use of ultrasound to detect the low level of antibodies in serum enhances the sensitivity of the test. The technique is based on the fact that suspended latex particles become concentrated in an ultrasonic standing wave field, thereby increasing the rate of particle-particle collisions compared to the standard agglutination test procedure. Reactions were performed on piezo-ceramic wafers, which were connected to an oscillator at a frequency of 100 kHz. This method improved sensitivity significantly so that even 20 times diluted serum samples exhibited agglutination, which was clearly visible to the naked eye.

Antibodies, Bacterial↗