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Application of direct agglutination test (DAT) and fast agglutination screening test (FAST) for sero-diagnosis of visceral leishmaniasis in endemic area of Minas Gerais, Brazil.

BACKGROUND: The direct agglutination test (DAT) has proved to be a very important sero-diagnostic tool combining high levels of intrinsic validity and ease of performance. Otherwise, fast agglutination screening test (FAST) utilises only one serum dilution making the test very suitable for the screening of large populations. RESULTS: We have tested FAST and DAT for the detection anti-Leishmania antibodies in serum samples from patients with American visceral (AVL) and cutaneous leishmaniases (ACL) in Minas Gerais State, Brazil. The DAT on serum and blood samples of confirmed AVL patients found all samples positive at a serum dilution of > or = 1:800. This dilution was subsequently used as cut off value in the study. The blood and serum samples of these confirmed patients could also be clearly read in FAST using a 1:100 dilution with the same high sensitivity. DAT and FAST were not able to detect significant amounts of antibodies in samples from ACL patients and are not suitable for the diagnosis of this manifestation of the disease. CONCLUSION: We suggest that both DAT and FAST are very practical diagnostic tools for the sero-diagnosis of AVL under rural conditions as both serological tests do not require sophisticated equipment, a cold chain and are very simple to perform.

Journal Article↗

Platelet agglutinating protein p37 causes platelet agglutination through its binding to membrane glycoprotein IV.

A 37 kDa platelet agglutinating protein (PAP p37) has previously been shown to be present in a subset of patients with thrombotic thrombocytopenic purpura and has been purified from their plasma. Using solubilized platelet membrane proteins from normal donors, it was shown by Western blotting that 125I-p37 bound to a membrane protein of 97 kDa (red/unred). Furthermore, the same protein was identified by reverse immunoblotting in which purified p37 was electrophoresed, transferred to the nitrocellulose sheet and incubated with solubilized normal platelet membrane proteins. The complex formed between p37 and the membrane protein was identified by autoradiography using polyclonal and monoclonal (OKM5) anti-GPIV antibodies, but was not detected by polyclonal antibody to GPIIIa. Similar studies with purified platelet GPIV under both reducing and non-reducing conditions demonstrated the binding of 125I-p37. Polyclonal and monoclonal antibodies to GPIV completely inhibited the platelet agglutination induced by TTP plasma containing p37, however, normal rabbit IgG, rabbit anti-GPIIIa IgG, and murine monoclonal anti-GPIIb/IIIa (10E5) antibodies had no effect. These data indicate that platelet GPIV is the receptor site for PAP p37.

Antibodies↗

Mating reaction in Saccharomyces cerevisiae. X. Agglutinability-inactivating factor: a factor which destroys sexual agglutinability of a mating-type cells.

From cells of Saccharomyces cerevisiae a factor has been extracted that destroys the agglutinability of a mating-type cells specifically. It was found in the cell extracts of diploid and tetraploid strains as well as haploid strains of a and alpha mating types. It is heat-labile and the molecular weight is about 50,000. It is adsorbed by neither a cells nor alpha cells. Its biological activity is dependent on the incubation temperature and the pH, and is completely inhibited by phenylmethylsulfonyl fluoride, a potent inhibitor of the serine proteases. All the results described in this paper indicate that this factor is a proteolytic enzyme.

Adsorption↗

Rapid identification of group A, B, C and G beta-haemolytic Streptococci by a modification of the co-agglutination technique. Comparison of results obtained by co-agglutination, fluorescent antibody test, counterimmunoelectrophoresis, and precipitin technique.

A rapid modification of the co-agglutination (COA) technique for grouping A, B, C and G beta-haemolytic streptococci was developed. The results are obtained within three hours after inoculation from primary plates of 0.5 ml broth. This method was compared to the fluorescent antibody test (FA) and counterimmunoelectrophoresis (CIE), two other rapid methods available for serological grouping of streptococci. Of 71 recently isolated streptococcal strains from clinical sources, 70 were correctly grouped in COA, 63 in FA and 68 in CIE. With commercial reagents COA compared favourably in accuracy to the other methods and can be recommended for routine serological grouping of beta-haemolytic streptococci in the clinical laboratory.

Agglutination Tests↗

Changes in the agglutinability of red cells and the inhibition of specific agglutination by plasma from human blood taken into ACD and stored at 4 degrees C.

The effect of storage in ACD at 4 degrees C on red cell agglutinability and the inhibitory properties of the plasma to blood group reagents has been studied. It was found that the variability demonstrated was related to the origin of the reagent used, particularly between human and on-human sources. The significance of these findings with regard to biochemical and morphological changes in blood on storage is discussed.

ABO Blood-Group System↗

[Methodical investigations to determine agglutination and inhibition of agglutination of cells from tumours and from thymus by the particle counter TuR ZG 2].

A report is given about a manyside applicable method to examine the agglutination of tumour cells and normal cells. It enables a presentation of results without subjective influences and is based on the electronic determination of the diminution of impulse numbers with a particle counter. This method is useful with little alterations for example to determine sugar specificity of lectins, effectivity of cytostatics and antisera against tumour cells and appropriate healthy cells.

Animals↗