Obtention and evaluation of Aspergillus fumigatus antigens extraction
Autor(a) principal: | |
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Data de Publicação: | 1992 |
Outros Autores: | , , |
Tipo de documento: | Artigo |
Idioma: | por |
Título da fonte: | Revista do Instituto de Medicina Tropical de São Paulo |
Texto Completo: | https://www.revistas.usp.br/rimtsp/article/view/28946 |
Resumo: | Antigens from three strains of Aspergillus fumigatus (354, 356, and JIG) and an antiserum against the mixing of these antigens have been produced, and evaluated immunochemically. The antigens were obtained through a modified Coleman & Kaufman technique (culture filtrate concentrated by acetone). Analysis by the immunodiffusion test (ID) against homologous serum has yielded 100% sensitivity (with the studied sera). Concerning heterologous sera we found reactivity with a serum of a patient of candidiasis and another with histoplasmosis. The same result was obtained with a reference antigen in immunodiffusion, showing similar standards of response. Titration of the antiserum by ID and counterimmunoelectrophoresis showed a title of 1:32, and by complement fixation (micro-technique) a title of 1:128. Using immunoelectrophoresis (IEF), the produced antiserum yielded 8 lines of precipitation (5 in the anodic pole and 3 in the cathodic one). In SDS-PAGE at 12.5% the antigen has presented a rather complex electro-phoretic profile (26 proteic subunits with a molecular weight ranging from 18 a >; 100 kDa). Immunogenicity of the antigen was observed in all fractions of SDS-PAGE when the immu-noblotting against the antiserum was carried out. |
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Obtention and evaluation of Aspergillus fumigatus antigens extraction Obtenção e avaliação de antígenos de Aspergillus fumigatus AntígenoAntissoroAspergillus fumigatus Antigens from three strains of Aspergillus fumigatus (354, 356, and JIG) and an antiserum against the mixing of these antigens have been produced, and evaluated immunochemically. The antigens were obtained through a modified Coleman & Kaufman technique (culture filtrate concentrated by acetone). Analysis by the immunodiffusion test (ID) against homologous serum has yielded 100% sensitivity (with the studied sera). Concerning heterologous sera we found reactivity with a serum of a patient of candidiasis and another with histoplasmosis. The same result was obtained with a reference antigen in immunodiffusion, showing similar standards of response. Titration of the antiserum by ID and counterimmunoelectrophoresis showed a title of 1:32, and by complement fixation (micro-technique) a title of 1:128. Using immunoelectrophoresis (IEF), the produced antiserum yielded 8 lines of precipitation (5 in the anodic pole and 3 in the cathodic one). In SDS-PAGE at 12.5% the antigen has presented a rather complex electro-phoretic profile (26 proteic subunits with a molecular weight ranging from 18 a >; 100 kDa). Immunogenicity of the antigen was observed in all fractions of SDS-PAGE when the immu-noblotting against the antiserum was carried out. Antígenos de três amostras de A. fumigatus (354, 356, JIG) e antissoro contra a mistura destes antígenos foram produzidos e avaliados imunoquimicamente. Os antígenos de filtrado de cultura foram obtidos após concentração com acetona conforme adaptação da técnica descrita por Coleman & Kaufman. Em prova de ID obteve-se 100% de positividade com os soros de pacientes com aspergilose estudados. Com relação aos soros heterólogos encontramos reatividade com soro de um paciente com candidíase e com soro de um paciente com histoplasmose; foi encontrado padrão idêntico de resposta quando se utilizou o antígeno de referência. O antissoro foi titulado por ID, CIE e RFC MI contra o antígeno específico apresentando títulos respectivos de 1:32, 1:32 e 1:128, e utilizado para reagir contra o mesmo antígeno por IEF, demonstrando 8 linhas de precipitação, sendo 5 na região anódica e 3 na região catódica. O perfil de bandeamento do antígeno em eletroforese utilizando gel de poliacrilamida (SDS-PAGE) a 12,5% apresentou-se complexo com 26 sub-unidades protéicas, cujos pesos moleculares variaram de 18 a >; 100kDa. Quando estes componentes foram eletrotransferidos e reagidos com o antissoro específico ("immunoblotting"), verificou-se imunogenicidade em todas as frações bandeadas. Universidade de São Paulo. Instituto de Medicina Tropical de São Paulo1992-08-01info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionapplication/pdfhttps://www.revistas.usp.br/rimtsp/article/view/28946Revista do Instituto de Medicina Tropical de São Paulo; Vol. 34 No. 4 (1992); 359-365 Revista do Instituto de Medicina Tropical de São Paulo; Vol. 34 Núm. 4 (1992); 359-365 Revista do Instituto de Medicina Tropical de São Paulo; v. 34 n. 4 (1992); 359-365 1678-99460036-4665reponame:Revista do Instituto de Medicina Tropical de São Pauloinstname:Instituto de Medicina Tropical (IMT)instacron:IMTporhttps://www.revistas.usp.br/rimtsp/article/view/28946/30803Copyright (c) 2018 Revista do Instituto de Medicina Tropical de São Pauloinfo:eu-repo/semantics/openAccessLirio, Vanda de SáAssis, Cezar Mendes deCano, Maria Isabel N.Lacaz, Carlos da Silva2012-07-02T01:33:23Zoai:revistas.usp.br:article/28946Revistahttp://www.revistas.usp.br/rimtsp/indexPUBhttps://www.revistas.usp.br/rimtsp/oai||revimtsp@usp.br1678-99460036-4665opendoar:2022-12-13T16:50:42.706969Revista do Instituto de Medicina Tropical de São Paulo - Instituto de Medicina Tropical (IMT)true |
dc.title.none.fl_str_mv |
Obtention and evaluation of Aspergillus fumigatus antigens extraction Obtenção e avaliação de antígenos de Aspergillus fumigatus |
title |
Obtention and evaluation of Aspergillus fumigatus antigens extraction |
spellingShingle |
Obtention and evaluation of Aspergillus fumigatus antigens extraction Lirio, Vanda de Sá Antígeno Antissoro Aspergillus fumigatus |
title_short |
Obtention and evaluation of Aspergillus fumigatus antigens extraction |
title_full |
Obtention and evaluation of Aspergillus fumigatus antigens extraction |
title_fullStr |
Obtention and evaluation of Aspergillus fumigatus antigens extraction |
title_full_unstemmed |
Obtention and evaluation of Aspergillus fumigatus antigens extraction |
title_sort |
Obtention and evaluation of Aspergillus fumigatus antigens extraction |
author |
Lirio, Vanda de Sá |
author_facet |
Lirio, Vanda de Sá Assis, Cezar Mendes de Cano, Maria Isabel N. Lacaz, Carlos da Silva |
author_role |
author |
author2 |
Assis, Cezar Mendes de Cano, Maria Isabel N. Lacaz, Carlos da Silva |
author2_role |
author author author |
dc.contributor.author.fl_str_mv |
Lirio, Vanda de Sá Assis, Cezar Mendes de Cano, Maria Isabel N. Lacaz, Carlos da Silva |
dc.subject.por.fl_str_mv |
Antígeno Antissoro Aspergillus fumigatus |
topic |
Antígeno Antissoro Aspergillus fumigatus |
description |
Antigens from three strains of Aspergillus fumigatus (354, 356, and JIG) and an antiserum against the mixing of these antigens have been produced, and evaluated immunochemically. The antigens were obtained through a modified Coleman & Kaufman technique (culture filtrate concentrated by acetone). Analysis by the immunodiffusion test (ID) against homologous serum has yielded 100% sensitivity (with the studied sera). Concerning heterologous sera we found reactivity with a serum of a patient of candidiasis and another with histoplasmosis. The same result was obtained with a reference antigen in immunodiffusion, showing similar standards of response. Titration of the antiserum by ID and counterimmunoelectrophoresis showed a title of 1:32, and by complement fixation (micro-technique) a title of 1:128. Using immunoelectrophoresis (IEF), the produced antiserum yielded 8 lines of precipitation (5 in the anodic pole and 3 in the cathodic one). In SDS-PAGE at 12.5% the antigen has presented a rather complex electro-phoretic profile (26 proteic subunits with a molecular weight ranging from 18 a >; 100 kDa). Immunogenicity of the antigen was observed in all fractions of SDS-PAGE when the immu-noblotting against the antiserum was carried out. |
publishDate |
1992 |
dc.date.none.fl_str_mv |
1992-08-01 |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/article info:eu-repo/semantics/publishedVersion |
format |
article |
status_str |
publishedVersion |
dc.identifier.uri.fl_str_mv |
https://www.revistas.usp.br/rimtsp/article/view/28946 |
url |
https://www.revistas.usp.br/rimtsp/article/view/28946 |
dc.language.iso.fl_str_mv |
por |
language |
por |
dc.relation.none.fl_str_mv |
https://www.revistas.usp.br/rimtsp/article/view/28946/30803 |
dc.rights.driver.fl_str_mv |
Copyright (c) 2018 Revista do Instituto de Medicina Tropical de São Paulo info:eu-repo/semantics/openAccess |
rights_invalid_str_mv |
Copyright (c) 2018 Revista do Instituto de Medicina Tropical de São Paulo |
eu_rights_str_mv |
openAccess |
dc.format.none.fl_str_mv |
application/pdf |
dc.publisher.none.fl_str_mv |
Universidade de São Paulo. Instituto de Medicina Tropical de São Paulo |
publisher.none.fl_str_mv |
Universidade de São Paulo. Instituto de Medicina Tropical de São Paulo |
dc.source.none.fl_str_mv |
Revista do Instituto de Medicina Tropical de São Paulo; Vol. 34 No. 4 (1992); 359-365 Revista do Instituto de Medicina Tropical de São Paulo; Vol. 34 Núm. 4 (1992); 359-365 Revista do Instituto de Medicina Tropical de São Paulo; v. 34 n. 4 (1992); 359-365 1678-9946 0036-4665 reponame:Revista do Instituto de Medicina Tropical de São Paulo instname:Instituto de Medicina Tropical (IMT) instacron:IMT |
instname_str |
Instituto de Medicina Tropical (IMT) |
instacron_str |
IMT |
institution |
IMT |
reponame_str |
Revista do Instituto de Medicina Tropical de São Paulo |
collection |
Revista do Instituto de Medicina Tropical de São Paulo |
repository.name.fl_str_mv |
Revista do Instituto de Medicina Tropical de São Paulo - Instituto de Medicina Tropical (IMT) |
repository.mail.fl_str_mv |
||revimtsp@usp.br |
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1798951639480008704 |