Entamoeba histolytica: detection of coproantigens by purified antibody in the capture sandwich ELISA

Detalhes bibliográficos
Autor(a) principal: Urdaneta, Haidee
Data de Publicação: 1994
Outros Autores: Guimarães, Semíramis [UNESP], Silva, Edward F., Tavares, Carlos A. P.
Tipo de documento: Artigo
Idioma: eng
Título da fonte: Repositório Institucional da UNESP
Texto Completo: http://dx.doi.org/10.1590/S0036-46651994000600011
http://hdl.handle.net/11449/211124
Resumo: A sensitive and specific Capture Sandwich ELISA (CSE) was developed using polyclonal purified rabbit antibodies against three different axenic strains of Entamoeba histolytica: CSP from Brazil and HM1 - IMSS from Mexico, for the detection of coproantigens in fecal samples. Immunoglobulin G (IgG) againstis E. histolytica was isolated from rabbits immunized with throphozoites whole extract in two stages: affinity chromatography in a column containing E. histolytica antigens bound to Sepharose 4B was followed by another chromatography in Sepharose antibodies 4B-Protein A. A Capture Sandwich ELISA using purified antibodies was able to detect 70ng of amebae protein, showing a sensitivity of 93% and specificity of 94%. The combination of microscopic examination and CSE gave a concordance and discordance of 93.25% and 6.75%, respectively. It was concluded that CSE is highly specific for the detection of coproantigens of E. histolytica in feces of infected patients, is quicker to perform, easier and more sensitive than microscopic examination.
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spelling Entamoeba histolytica: detection of coproantigens by purified antibody in the capture sandwich ELISAEntamoeba histolytica: detecção de coproantígenos por ELISA de captura utilizando anticorpo purificadoEntamoeba histolyticaAmebiasisCoproantigensImmunodiagnosticPurified antibodyCapture Sandwich ELISA (CSE)A sensitive and specific Capture Sandwich ELISA (CSE) was developed using polyclonal purified rabbit antibodies against three different axenic strains of Entamoeba histolytica: CSP from Brazil and HM1 - IMSS from Mexico, for the detection of coproantigens in fecal samples. Immunoglobulin G (IgG) againstis E. histolytica was isolated from rabbits immunized with throphozoites whole extract in two stages: affinity chromatography in a column containing E. histolytica antigens bound to Sepharose 4B was followed by another chromatography in Sepharose antibodies 4B-Protein A. A Capture Sandwich ELISA using purified antibodies was able to detect 70ng of amebae protein, showing a sensitivity of 93% and specificity of 94%. The combination of microscopic examination and CSE gave a concordance and discordance of 93.25% and 6.75%, respectively. It was concluded that CSE is highly specific for the detection of coproantigens of E. histolytica in feces of infected patients, is quicker to perform, easier and more sensitive than microscopic examination.Foi desenvolvido um teste de ELISA de Captura usando anticorpos policlonais purificados obtidos em coelhos contra três diferentes cepas axênicas de Entamoeba histolytica (ICB-CSP and ICB-462 do Brasil e HM1 do México) para detecção de coproantígenos em amostras de fezes de indivíduos: a) sintomáticos, b) assintomáticos, c) com outros parasitos intestinais, e d) sadios. Imunoglobulina G (IgG) contra E. histolytica foi isolada de imune soro de coelho, em duas etapas: cromatografia de afinidade em uma coluna contendo antígenos de E. histolytica unidos à Sepharose 4B, seguido por outra cromatografía em Sepharose 4B Proteína A. O teste de ELISA usando anticorpos purificados, foi capaz de detectar até um só trofozoíto por lâmina ou 70 ng de proteína de ameba por orifício, apresentando uma sensibilidade de 93% e uma especificidade de 94%. A combinação do exame microscópico com o método de ELISA de Captura teve uma concordância e discordância de 93,25% e 6,75%, respectivamente. Pode-se concluir que o teste de ELISA de Captura utilizando anticorpo purificado é altamente específico para a detecção de coproantígenos de E. histolytica em fezes de pacientes infectados, rápido, fácil e mais sensível que o exame microscópico.Department of Biochemistry and ImmunologyUniversidade Federal de Minas Gerais, Institute of Biological SciencesLos Andes University, Department of BiologyUniversidade Estadual Paulista, Biociences InstituteUniversidade Estadual Paulista, Biociences InstituteInstituto de Medicina TropicalDepartment of Biochemistry and ImmunologyUniversidade Federal de Minas GeraisLos Andes UniversityUniversidade Estadual Paulista (Unesp)Urdaneta, HaideeGuimarães, Semíramis [UNESP]Silva, Edward F.Tavares, Carlos A. P.2021-07-14T10:19:38Z2021-07-14T10:19:38Z1994-12info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/article539-545application/pdfhttp://dx.doi.org/10.1590/S0036-46651994000600011Revista do Instituto de Medicina Tropical de São Paulo. São Paulo, SP, Brazil: Instituto de Medicina Tropical, v. 36, n. 6, p. 539-545, 1994.1678-9946http://hdl.handle.net/11449/21112410.1590/S0036-46651994000600011S0036-46651994000600011S0036-46651994000600011.pdfSciELOreponame:Repositório Institucional da UNESPinstname:Universidade Estadual Paulista (UNESP)instacron:UNESPengRevista do Instituto de Medicina Tropical de São Pauloinfo:eu-repo/semantics/openAccess2023-11-08T06:13:47Zoai:repositorio.unesp.br:11449/211124Repositório InstitucionalPUBhttp://repositorio.unesp.br/oai/requestopendoar:29462023-11-08T06:13:47Repositório Institucional da UNESP - Universidade Estadual Paulista (UNESP)false
dc.title.none.fl_str_mv Entamoeba histolytica: detection of coproantigens by purified antibody in the capture sandwich ELISA
Entamoeba histolytica: detecção de coproantígenos por ELISA de captura utilizando anticorpo purificado
title Entamoeba histolytica: detection of coproantigens by purified antibody in the capture sandwich ELISA
spellingShingle Entamoeba histolytica: detection of coproantigens by purified antibody in the capture sandwich ELISA
Urdaneta, Haidee
Entamoeba histolytica
Amebiasis
Coproantigens
Immunodiagnostic
Purified antibody
Capture Sandwich ELISA (CSE)
title_short Entamoeba histolytica: detection of coproantigens by purified antibody in the capture sandwich ELISA
title_full Entamoeba histolytica: detection of coproantigens by purified antibody in the capture sandwich ELISA
title_fullStr Entamoeba histolytica: detection of coproantigens by purified antibody in the capture sandwich ELISA
title_full_unstemmed Entamoeba histolytica: detection of coproantigens by purified antibody in the capture sandwich ELISA
title_sort Entamoeba histolytica: detection of coproantigens by purified antibody in the capture sandwich ELISA
author Urdaneta, Haidee
author_facet Urdaneta, Haidee
Guimarães, Semíramis [UNESP]
Silva, Edward F.
Tavares, Carlos A. P.
author_role author
author2 Guimarães, Semíramis [UNESP]
Silva, Edward F.
Tavares, Carlos A. P.
author2_role author
author
author
dc.contributor.none.fl_str_mv Department of Biochemistry and Immunology
Universidade Federal de Minas Gerais
Los Andes University
Universidade Estadual Paulista (Unesp)
dc.contributor.author.fl_str_mv Urdaneta, Haidee
Guimarães, Semíramis [UNESP]
Silva, Edward F.
Tavares, Carlos A. P.
dc.subject.por.fl_str_mv Entamoeba histolytica
Amebiasis
Coproantigens
Immunodiagnostic
Purified antibody
Capture Sandwich ELISA (CSE)
topic Entamoeba histolytica
Amebiasis
Coproantigens
Immunodiagnostic
Purified antibody
Capture Sandwich ELISA (CSE)
description A sensitive and specific Capture Sandwich ELISA (CSE) was developed using polyclonal purified rabbit antibodies against three different axenic strains of Entamoeba histolytica: CSP from Brazil and HM1 - IMSS from Mexico, for the detection of coproantigens in fecal samples. Immunoglobulin G (IgG) againstis E. histolytica was isolated from rabbits immunized with throphozoites whole extract in two stages: affinity chromatography in a column containing E. histolytica antigens bound to Sepharose 4B was followed by another chromatography in Sepharose antibodies 4B-Protein A. A Capture Sandwich ELISA using purified antibodies was able to detect 70ng of amebae protein, showing a sensitivity of 93% and specificity of 94%. The combination of microscopic examination and CSE gave a concordance and discordance of 93.25% and 6.75%, respectively. It was concluded that CSE is highly specific for the detection of coproantigens of E. histolytica in feces of infected patients, is quicker to perform, easier and more sensitive than microscopic examination.
publishDate 1994
dc.date.none.fl_str_mv 1994-12
2021-07-14T10:19:38Z
2021-07-14T10:19:38Z
dc.type.status.fl_str_mv info:eu-repo/semantics/publishedVersion
dc.type.driver.fl_str_mv info:eu-repo/semantics/article
format article
status_str publishedVersion
dc.identifier.uri.fl_str_mv http://dx.doi.org/10.1590/S0036-46651994000600011
Revista do Instituto de Medicina Tropical de São Paulo. São Paulo, SP, Brazil: Instituto de Medicina Tropical, v. 36, n. 6, p. 539-545, 1994.
1678-9946
http://hdl.handle.net/11449/211124
10.1590/S0036-46651994000600011
S0036-46651994000600011
S0036-46651994000600011.pdf
url http://dx.doi.org/10.1590/S0036-46651994000600011
http://hdl.handle.net/11449/211124
identifier_str_mv Revista do Instituto de Medicina Tropical de São Paulo. São Paulo, SP, Brazil: Instituto de Medicina Tropical, v. 36, n. 6, p. 539-545, 1994.
1678-9946
10.1590/S0036-46651994000600011
S0036-46651994000600011
S0036-46651994000600011.pdf
dc.language.iso.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv Revista do Instituto de Medicina Tropical de São Paulo
dc.rights.driver.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv 539-545
application/pdf
dc.publisher.none.fl_str_mv Instituto de Medicina Tropical
publisher.none.fl_str_mv Instituto de Medicina Tropical
dc.source.none.fl_str_mv SciELO
reponame:Repositório Institucional da UNESP
instname:Universidade Estadual Paulista (UNESP)
instacron:UNESP
instname_str Universidade Estadual Paulista (UNESP)
instacron_str UNESP
institution UNESP
reponame_str Repositório Institucional da UNESP
collection Repositório Institucional da UNESP
repository.name.fl_str_mv Repositório Institucional da UNESP - Universidade Estadual Paulista (UNESP)
repository.mail.fl_str_mv
_version_ 1803046384624992256

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