Obtention and evaluation of Aspergillus fumigatus antigens extraction

Detalhes bibliográficos
Autor(a) principal: Lirio, Vanda de Sá
Data de Publicação: 1992
Outros Autores: Assis, Cezar Mendes de, Cano, Maria Isabel N., Lacaz, Carlos da Silva
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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spelling 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)
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