Detection of giardia duodenalis antigen in human fecal eluates by enzyme-linked immunosorbent assay using polyclonal antibodies

The present study developed and standardized an enzime-linked immunosorbent assay (ELISA) to detect Giardia antigen in feces using rabbit polyclonal antibodies. Giardia cysts were purified from human fecal samples by sucrose and percoll gradients. Gerbils (Meriones unguiculatus) were infected to obt...

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Autores:
Duque Beltrán, Sofía
Nicholls Orejuela, Rubén Santiago
Arévalo Jamaica, Adriana
Guerrero-Lozano, Rafael
Montenegro, Sonia
James, Mark A
Tipo de recurso:
Fecha de publicación:
2002
Institución:
Universidad El Bosque
Repositorio:
Repositorio U. El Bosque
Idioma:
eng
OAI Identifier:
oai:repositorio.unbosque.edu.co:20.500.12495/1661
Acceso en línea:
http://hdl.handle.net/20.500.12495/1661
http://doi.org/10.1590/S0074-02762002000800018
Palabra clave:
Ensayo de inmunoadsorción enzimática
Formación de anticuerpos
Pruebas inmunológicas
Giardia
Feces
Antigen
Rights
License
Attribution-NonCommercial 4.0 International
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network_acronym_str UNBOSQUE2
network_name_str Repositorio U. El Bosque
repository_id_str
dc.title.spa.fl_str_mv Detection of giardia duodenalis antigen in human fecal eluates by enzyme-linked immunosorbent assay using polyclonal antibodies
title Detection of giardia duodenalis antigen in human fecal eluates by enzyme-linked immunosorbent assay using polyclonal antibodies
spellingShingle Detection of giardia duodenalis antigen in human fecal eluates by enzyme-linked immunosorbent assay using polyclonal antibodies
Ensayo de inmunoadsorción enzimática
Formación de anticuerpos
Pruebas inmunológicas
Giardia
Feces
Antigen
title_short Detection of giardia duodenalis antigen in human fecal eluates by enzyme-linked immunosorbent assay using polyclonal antibodies
title_full Detection of giardia duodenalis antigen in human fecal eluates by enzyme-linked immunosorbent assay using polyclonal antibodies
title_fullStr Detection of giardia duodenalis antigen in human fecal eluates by enzyme-linked immunosorbent assay using polyclonal antibodies
title_full_unstemmed Detection of giardia duodenalis antigen in human fecal eluates by enzyme-linked immunosorbent assay using polyclonal antibodies
title_sort Detection of giardia duodenalis antigen in human fecal eluates by enzyme-linked immunosorbent assay using polyclonal antibodies
dc.creator.fl_str_mv Duque Beltrán, Sofía
Nicholls Orejuela, Rubén Santiago
Arévalo Jamaica, Adriana
Guerrero-Lozano, Rafael
Montenegro, Sonia
James, Mark A
dc.contributor.author.none.fl_str_mv Duque Beltrán, Sofía
Nicholls Orejuela, Rubén Santiago
Arévalo Jamaica, Adriana
Guerrero-Lozano, Rafael
Montenegro, Sonia
James, Mark A
dc.subject.decs.spa.fl_str_mv Ensayo de inmunoadsorción enzimática
Formación de anticuerpos
Pruebas inmunológicas
topic Ensayo de inmunoadsorción enzimática
Formación de anticuerpos
Pruebas inmunológicas
Giardia
Feces
Antigen
dc.subject.keywords.spa.fl_str_mv Giardia
Feces
Antigen
description The present study developed and standardized an enzime-linked immunosorbent assay (ELISA) to detect Giardia antigen in feces using rabbit polyclonal antibodies. Giardia cysts were purified from human fecal samples by sucrose and percoll gradients. Gerbils (Meriones unguiculatus) were infected to obtain trophozoites. Rabbits were inoculated with either cyst or trophozoite antigens of 14 Colombian Giardia isolates to develop antibodies against the respective stages. The IgG anti-Giardia were purified by sequential caprylic acid and ammonium sulfate precipitation. A portion of these polyclonal antibodies was linked to alkaline phosphatase (conjugate). One hundred and ninety six samples of human feces, from different patients, were tested by parasitologic diagnosis: 69 were positive for Giardia cysts, 56 had no Giardia parasites, and 71 revealed parasites other than Giardia. The optimal concentration of polyclonal antibodies for antigen capture was 40 µg/ml and the optimal conjugate dilution was 1:100. The absorbance cut-off value was 0.24. The parameters of the ELISA test for Giardia antigen detection were: sensitivity, 100% (95% CI: 93.4-100%); specificity, 95% (95% CI: 88.6-97.6%); positive predictive value, 91% (95% CI: 81.4-95.9%); and negative predictive value, 100% (95% CI: 96.1-100%). This ELISA will improve the diagnosis of Giardia infections in Colombia and will be useful in following patients after treatment.
publishDate 2002
dc.date.issued.none.fl_str_mv 2002
dc.date.accessioned.none.fl_str_mv 2019-08-27T16:40:13Z
dc.date.available.none.fl_str_mv 2019-08-27T16:40:13Z
dc.type.spa.fl_str_mv article
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dc.type.local.spa.fl_str_mv artículo
dc.identifier.issn.none.fl_str_mv 1678-8060
dc.identifier.uri.none.fl_str_mv http://hdl.handle.net/20.500.12495/1661
dc.identifier.doi.none.fl_str_mv http://doi.org/10.1590/S0074-02762002000800018
dc.identifier.instname.spa.fl_str_mv instname:Universidad El Bosque
dc.identifier.reponame.spa.fl_str_mv reponame:Repositorio Institucional Universidad El Bosque
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identifier_str_mv 1678-8060
instname:Universidad El Bosque
reponame:Repositorio Institucional Universidad El Bosque
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url http://hdl.handle.net/20.500.12495/1661
http://doi.org/10.1590/S0074-02762002000800018
dc.language.iso.none.fl_str_mv eng
language eng
dc.relation.ispartofseries.spa.fl_str_mv Memorias do Instituto Oswaldo Cruz, 1678-8060, Vol, 97, Nro. 8, 2002, p. 1165-1168
dc.relation.uri.none.fl_str_mv http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0074-02762002000800018&lng=en&tlng=en
dc.rights.*.fl_str_mv Attribution-NonCommercial 4.0 International
dc.rights.coar.fl_str_mv http://purl.org/coar/access_right/c_abf2
dc.rights.uri.*.fl_str_mv http://creativecommons.org/licenses/by-nc/4.0/
dc.rights.local.spa.fl_str_mv Acceso abierto
dc.rights.accessrights.none.fl_str_mv http://purl.org/coar/access_right/c_abf125
dc.rights.creativecommons.none.fl_str_mv 2002
rights_invalid_str_mv Attribution-NonCommercial 4.0 International
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Acceso abierto
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http://purl.org/coar/access_right/c_abf2
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dc.publisher.spa.fl_str_mv Instituto Oswaldo Cruz, Ministério da Saúde
dc.publisher.journal.spa.fl_str_mv Memorias do Instituto Oswaldo Cruz
institution Universidad El Bosque
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spelling Duque Beltrán, SofíaNicholls Orejuela, Rubén SantiagoArévalo Jamaica, AdrianaGuerrero-Lozano, RafaelMontenegro, SoniaJames, Mark A2019-08-27T16:40:13Z2019-08-27T16:40:13Z20021678-8060http://hdl.handle.net/20.500.12495/1661http://doi.org/10.1590/S0074-02762002000800018instname:Universidad El Bosquereponame:Repositorio Institucional Universidad El Bosquerepourl:https://repositorio.unbosque.edu.coapplication/pdfengInstituto Oswaldo Cruz, Ministério da SaúdeMemorias do Instituto Oswaldo CruzMemorias do Instituto Oswaldo Cruz, 1678-8060, Vol, 97, Nro. 8, 2002, p. 1165-1168http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0074-02762002000800018&lng=en&tlng=enAttribution-NonCommercial 4.0 Internationalhttp://creativecommons.org/licenses/by-nc/4.0/Acceso abiertohttp://purl.org/coar/access_right/c_abf1252002http://purl.org/coar/access_right/c_abf2Detection of giardia duodenalis antigen in human fecal eluates by enzyme-linked immunosorbent assay using polyclonal antibodiesarticleartículohttp://purl.org/coar/version/c_970fb48d4fbd8a85http://purl.org/coar/resource_type/c_6501Ensayo de inmunoadsorción enzimáticaFormación de anticuerposPruebas inmunológicasGiardiaFecesAntigenThe present study developed and standardized an enzime-linked immunosorbent assay (ELISA) to detect Giardia antigen in feces using rabbit polyclonal antibodies. Giardia cysts were purified from human fecal samples by sucrose and percoll gradients. Gerbils (Meriones unguiculatus) were infected to obtain trophozoites. Rabbits were inoculated with either cyst or trophozoite antigens of 14 Colombian Giardia isolates to develop antibodies against the respective stages. The IgG anti-Giardia were purified by sequential caprylic acid and ammonium sulfate precipitation. A portion of these polyclonal antibodies was linked to alkaline phosphatase (conjugate). One hundred and ninety six samples of human feces, from different patients, were tested by parasitologic diagnosis: 69 were positive for Giardia cysts, 56 had no Giardia parasites, and 71 revealed parasites other than Giardia. The optimal concentration of polyclonal antibodies for antigen capture was 40 µg/ml and the optimal conjugate dilution was 1:100. The absorbance cut-off value was 0.24. The parameters of the ELISA test for Giardia antigen detection were: sensitivity, 100% (95% CI: 93.4-100%); specificity, 95% (95% CI: 88.6-97.6%); positive predictive value, 91% (95% CI: 81.4-95.9%); and negative predictive value, 100% (95% CI: 96.1-100%). 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