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1.
Bioorg Khim ; 20(7): 731-9, 1994 Jul.
Article in Russian | MEDLINE | ID: mdl-7993377

ABSTRACT

Non-instrumental immunoassays based on immunofiltration and microtiter particle agglutination (MPA) techniques have been developed using coloured polyacrolein latex. These methods have been applied to the quantification of the group-specific polysaccharide (PS) of S. pyogenes (group A streptococcus) and compared with standard ELISA tests. The most efficient method was MPA; as little as 0.05 ng A-PS/ml could be detected in 1.5 h. In comparison with ELISA test, the sensitivity of MPA was 10 times higher and the procedure was much simpler. The sensitivity of immunofiltration assay using both enzyme and latex conjugates was shown to be the same (50 ng/ml A-PS) and the duration of the assay 3-5 min. No cross-reactions of latex conjugates with non A streptococcus cell lysates have been observed. The developed methods are rapid, robust, easy to perform, don't need any sophisticated equipment and specially trained staff.


Subject(s)
Acrolein , Latex , Polymers , Polysaccharides, Bacterial/analysis , Streptococcus pyogenes/chemistry , Coloring Agents , Filtration , Immunoenzyme Techniques
2.
Zh Mikrobiol Epidemiol Immunobiol ; (11): 69-71, 1988 Nov.
Article in Russian | MEDLINE | ID: mdl-2464262

ABSTRACT

O-specific polysaccharide (PS) obtained from P. aeruginosa lipopolysaccharide (LPS), immunotypes 1, 2 and 7, were condensed in the presence of aminated bovine serum albumin (amBSA). As a result, artificial polysaccharide-protein antigens were synthesized and their protective properties were studied by the subcutaneous immunization of mice with their subsequent challenge with the cells of the homologous strain injected intraperitoneally in a dose of 3-5 LD50. The most effective immunization was achieved by using homologous LPS, taken in a subimmunogenic dose, as adjuvant. Heterologous LPS, taken in the same doses, showed no stimulating effect. The protective properties of the conjugates were also found to depend on the length of the PS chain. The conjugate of amBSA and PS of immunotype 2 with a molecular weight of about 10 KD was found to possess the highest protective potency. The decrease of the molecular weight of PS of immunotype 7 from 50 KD to 10 KD by selective acid hydrolysis led to obtaining the conjugate with high protective potency. The possibility of using the structurally linked pair antigen-adjuvant for the development of artificial vaccines is discussed.


Subject(s)
Antigens, Bacterial/immunology , Bacterial Vaccines/immunology , Epitopes/immunology , Polysaccharides/immunology , Pseudomonas aeruginosa/immunology , Animals , Drug Evaluation, Preclinical , Exotoxins/immunology , Immunization , Mice , Molecular Weight , O Antigens , Pseudomonas Infections/prevention & control , Serum Albumin, Bovine/immunology , Vaccines, Synthetic/immunology
4.
Mol Gen Mikrobiol Virusol ; (5): 44-6, 1987 May.
Article in Russian | MEDLINE | ID: mdl-3112564

ABSTRACT

A modification of the traditional method for lipoplysaccharide isolation from the cells of grammnegative bacteria was elaborated on the basis of extraction by the hot water solution of phenol (the method of Westfahl). To make the method simpler and to raise the yield of the product it was proposed to use the water-phenol extract without its division for plases. The nucleic acids are eliminated by precipitation from dialyzed extract at pH 3,2-3,4 achieved by addition of acetic acid. The comparative isolation of lipopolysaccharides by the classic and modified methods has confirmed the advantages of a new technique.


Subject(s)
Gram-Negative Bacteria/analysis , Lipopolysaccharides/isolation & purification , Gram-Negative Bacteria/immunology , Methods , Pseudomonas aeruginosa/analysis , Pseudomonas aeruginosa/immunology , Shigella boydii/analysis , Shigella boydii/immunology
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