RESUMO
The results obtained in the study of the possibility of using magnetic sorbents for the construction of a diagnostic assay system based on the antigen-antibody interaction are presented. As a model, Yersinia pestis capsular antigen and immunoglobulins to it have been used. A solid-phase immunofluorescent liposomal assay method has been developed; this method can be used for the detection of biopolymers in the sample under study and for the determination of their activity.
Assuntos
Testes Imunológicos/métodos , Imunoadsorventes , Lipossomos , Magnetismo , Resinas Acrílicas , Animais , Reações Antígeno-Anticorpo/imunologia , Antígenos de Bactérias/análise , Biopolímeros , Imunização , Imunoglobulinas/análise , Testes Imunológicos/instrumentação , Peste/diagnóstico , Coelhos , Yersinia pestis/imunologiaAssuntos
Técnicas de Imunoadsorção , Resinas Acrílicas , Gangliosídeos , Humanos , Imunoglobulinas , SefaroseRESUMO
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.
Assuntos
Enterotoxinas/análise , Vibrio cholerae/análise , ImunofluorescênciaRESUMO
In this work the possibility of using neuraminidase for increasing the content of ganglioside GM1 in the mixture of gangliosides used for the sensitization of erythrocytes has been studied. The study has revealed that the treatment of gangliosides with neuraminidase is sufficient for obtaining active hemosensitin; there is no need for the purification of the preparation by gel filtration.
Assuntos
Toxina da Cólera/isolamento & purificação , Eritrócitos , Gangliosídeos , Neuraminidase , Vibrio cholerae/enzimologia , Animais , Cromatografia em Camada Fina , Testes de Hemaglutinação , Ovinos , SuínosRESUMO
The studies described in this work have indicated that cholera enterotoxin and its components (cholerogenoid and subunit B) can be detected in amounts of 0.25, 0.28 and 0.6 microgram of protein per 1 ml, respectively, by means of erythrocytes sensitized with gangliozide-containing complex. The conditions for erythrocyte sensitization have been established. The methods of cholerogen titration in the passive hemagglutination test by means of the erythrocytic gangliozide diagnostic reagent and in Craig's skin test have been shown to correlate. Similarly, the passive hemagglutination test is supposed to be suitable for detecting other bacterial toxins interacting with gangliozides.
Assuntos
Toxina da Cólera/análise , Enterotoxinas/análise , Toxoides/análise , Vibrio cholerae/análise , Animais , Química Encefálica , Eritrócitos/imunologia , Gangliosídeos/isolamento & purificação , Testes de Hemaglutinação/métodos , Testes Cutâneos , SuínosRESUMO
The possibility of obtaining biologically active cholerogen by extracting it from solid culture media used for the cultivation of vibrios is shown. V. cholerae, strain 569B, cultivated on solid media produced about 3 times more toxin per 1 ml of the culture medium than during the process of its cultivation with the use of liquid culture media. The inculation temperature proved to have no essential influence on the toxin production by vibrios.