Asunto(s)
Complejo Antígeno-Anticuerpo/aislamiento & purificación , Técnicas de Inmunoadsorción , Inmunoadsorbentes/aislamiento & purificación , Magnetismo , Técnica del Anticuerpo Fluorescente/instrumentación , Técnicas de Inmunoadsorción/instrumentación , Ligandos , Magnetismo/instrumentación , Técnicas Microbiológicas/instrumentación , Polímeros , PolvosRESUMEN
The detection of Y. pestis cells in magnetic enzyme immunoassay is carried out with the use of magnetic polyacrylamide microgranules. In the assay system for the determination of the antigen commercial Y. pestis antigens, peroxidase-labeled antibodies, the substrate mixture consisting of sodium salt of 2,2-azino-bis-3-ethylbenzthiazoline-6-sulfonic acid and H2O2 in citrate-phosphate buffer solution, pH 4.5, are used. The sensitivity of the method is 5 X 10(4) microbial bodies per ml.
Asunto(s)
Antígenos Bacterianos/análisis , Técnicas para Inmunoenzimas , Magnetismo/métodos , Yersinia pestis/inmunología , Adsorción , Estudios de Evaluación como Asunto , Técnicas para Inmunoenzimas/normas , Inmunoadsorbentes , Magnetismo/normas , SolubilidadRESUMEN
Antisera to the antigens of 5 fractions, isolated as the result of the separation of P. pseudomallei aqueous saline extract by continuous electrophoresis in the vertical block of granulated gel, have been obtained. Immunoelectrophoresis with the use of P. pseudomallei aqueous saline extract has revealed that antisera to electrophoretic fractions contain antibodies mainly to the antigens of the corresponding fractions, which shows that this technique ensures the effective separation of P. pseudomallei biopolymers by their electrophoretic motility and molecular weight. These antisera differ in their species specificity. Thus, antisera to antigens with anode motility have been found to contain antibodies mainly to P. pseudomallei antigens and antisera to electroneutral antigens or to those with cathode motility, to P. pseudomallei and P. mallei antigens.
Asunto(s)
Antígenos Bacterianos/inmunología , Melioidosis/diagnóstico , Pseudomonas/inmunología , Animales , Anticuerpos Antibacterianos/análisis , Antígenos Bacterianos/aislamiento & purificación , Relación Dosis-Respuesta Inmunológica , Electroforesis en Gel Bidimensional , Técnica del Anticuerpo Fluorescente , Inmunización , Inmunodifusión , ConejosRESUMEN
The 96% ethanol has been found fairly effective for the fixation and disinfection of Yersinia pestis, Vibrio cholerae, Pseudomonas mallei, and Pseudomonas pseudomallei after their contact with magnetic polyacrylamide sorbents containing specific immunoglobulins for ligands. Such step is recommended for the quantitative immunofluorescence test employed for rapid analysis, diagnosis, and identification of the above microorganisms.
Asunto(s)
Técnicas Bacteriológicas , Técnica del Anticuerpo FluorescenteRESUMEN
The adsorption capacity of microgranulated polyacrylamide magnetic immunosorbents has been studied by the method of quantitative immunofluorescence as applied to the causative agents of plague, cholera, and melioidosis. Similar regularity in the dynamics of antigen adsorption on the granules of magnetic immunoadsorbents has been established. This regularity consists in the direct relationship between this process and the concentration of infective agents interacting with magnetic sorbents.
Asunto(s)
Complejo Antígeno-Anticuerpo/farmacología , Técnica del Anticuerpo Fluorescente , Inmunoadsorbentes/farmacología , Magnetismo , Resinas Acrílicas/farmacología , Adsorción , Antígenos Bacterianos/análisis , Técnicas de Inmunoadsorción , Pseudomonas/inmunología , Factores de Tiempo , Vibrio cholerae/inmunología , Yersinia pestis/inmunologíaAsunto(s)
Técnicas de Inmunoadsorción , Resinas Acrílicas , Gangliósidos , Humanos , Inmunoglobulinas , SefarosaRESUMEN
A variant of EIA techniques for the determination of cholera enterotoxin is proposed. This method is based on the selective sorption of the toxin on ganglioside-containing magnetic granules with its subsequent detection by means of immune serum and antispecific immunoperoxidase conjugate. The proposed method permits the detection of 0.052 +/- 0.02 ng of protein of the purified toxin.
Asunto(s)
Enterotoxinas/análisis , Gangliósidos , Magnetismo , Vibrio cholerae , Resinas Acrílicas , Toxina del Cólera/análisis , Ensayo de Inmunoadsorción Enzimática , Geles , Técnicas para Inmunoenzimas/instrumentación , Inmunoadsorbentes , Pruebas CutáneasRESUMEN
The quantitative immunofluorescent assay for the determination of cholera enterotoxin is proposed. The assay is based on the selective sorption of cholera enterotoxin by gangliosides incorporated into polyacrylamide granules. The preliminary treatment of gangliosides with neuraminidase enhances the sensitivity of this assay. The assay permits the detection of cholerigen in an amount of 20 ng.
Asunto(s)
Enterotoxinas/análisis , Vibrio cholerae/análisis , Técnica del Anticuerpo FluorescenteRESUMEN
In this work simple techniques for obtaining polyacrylamide sorbents with magnetic properties are described. These techniques have permitted obtaining block and microgranulated sorbents with the immobilization of antibodies from plague antiserum in the cellular gel structure for the specific sorption of killed and live Yersinia pestis cells and their first fraction; pig brain gangliosides have also been incorporated into the gel structure with a view to the sorption of cholera toxin from the filtrate of Vibrio cholerae culture. The magnetic properties of sorbents, obtained by the copolymerization of powdered magnetic ferric oxides in gel, have made it possible to increase the effectiveness of specific sorption due to mixing and rapid separation in different magnetic fields, as well as to facilitate and accelerate manipulations with the sorbent at all stages. The capacity of different types of sorbents and the time of sorption have been determined.