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...
- 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
id |
UNBOSQUE2_4d83eafab8af3609f54838a4d9281123 |
---|---|
oai_identifier_str |
oai:repositorio.unbosque.edu.co:20.500.12495/1661 |
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 |
dc.type.coarversion.fl_str_mv |
http://purl.org/coar/version/c_970fb48d4fbd8a85 |
dc.type.coar.fl_str_mv |
http://purl.org/coar/resource_type/c_6501 |
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 |
dc.identifier.repourl.none.fl_str_mv |
repourl:https://repositorio.unbosque.edu.co |
identifier_str_mv |
1678-8060 instname:Universidad El Bosque reponame:Repositorio Institucional Universidad El Bosque repourl:https://repositorio.unbosque.edu.co |
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 http://creativecommons.org/licenses/by-nc/4.0/ Acceso abierto http://purl.org/coar/access_right/c_abf125 2002 http://purl.org/coar/access_right/c_abf2 |
dc.format.mimetype.none.fl_str_mv |
application/pdf |
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 |
bitstream.url.fl_str_mv |
https://repositorio.unbosque.edu.co/bitstreams/ee122ac2-7bca-42ad-990a-2e48b852baba/download https://repositorio.unbosque.edu.co/bitstreams/72349d43-25ee-46d5-a080-cf1a097b98a5/download https://repositorio.unbosque.edu.co/bitstreams/081c3b74-f887-4f45-988d-484c9a6a88d8/download https://repositorio.unbosque.edu.co/bitstreams/116a7ac3-8b29-4166-8a24-f381018d221b/download https://repositorio.unbosque.edu.co/bitstreams/d63d2dac-cf76-40c6-814d-621b7535cdca/download |
bitstream.checksum.fl_str_mv |
584af5cf662ad67960ac08995f59aec2 24013099e9e6abb1575dc6ce0855efd5 8a4605be74aa9ea9d79846c1fba20a33 f1c39dd38c283f91f5987b58daab3868 a83940a80fa7a1b61e800ce4958ae01e |
bitstream.checksumAlgorithm.fl_str_mv |
MD5 MD5 MD5 MD5 MD5 |
repository.name.fl_str_mv |
Repositorio Institucional Universidad El Bosque |
repository.mail.fl_str_mv |
bibliotecas@biteca.com |
_version_ |
1814100783959900160 |
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%). This ELISA will improve the diagnosis of Giardia infections in Colombia and will be useful in following patients after treatment.ORIGINALDuque-Beltrán S., Nicholls-Orejuela R.S., Arévalo-Jamaica A., Guerrero-Lozano R., Montenegro S., James M.A._2002.pdfDuque-Beltrán S., Nicholls-Orejuela R.S., Arévalo-Jamaica A., Guerrero-Lozano R., Montenegro S., James M.A._2002.pdfapplication/pdf57060https://repositorio.unbosque.edu.co/bitstreams/ee122ac2-7bca-42ad-990a-2e48b852baba/download584af5cf662ad67960ac08995f59aec2MD51CC-LICENSElicense_rdflicense_rdfapplication/rdf+xml; charset=utf-8914https://repositorio.unbosque.edu.co/bitstreams/72349d43-25ee-46d5-a080-cf1a097b98a5/download24013099e9e6abb1575dc6ce0855efd5MD52LICENSElicense.txtlicense.txttext/plain; charset=utf-81748https://repositorio.unbosque.edu.co/bitstreams/081c3b74-f887-4f45-988d-484c9a6a88d8/download8a4605be74aa9ea9d79846c1fba20a33MD53THUMBNAILDuque-Beltrán S., Nicholls-Orejuela R.S., Arévalo-Jamaica A., Guerrero-Lozano R., Montenegro S., James M.A._2002.pdf.jpgDuque-Beltrán S., Nicholls-Orejuela R.S., Arévalo-Jamaica A., Guerrero-Lozano R., Montenegro S., James M.A._2002.pdf.jpgIM Thumbnailimage/jpeg8990https://repositorio.unbosque.edu.co/bitstreams/116a7ac3-8b29-4166-8a24-f381018d221b/downloadf1c39dd38c283f91f5987b58daab3868MD54TEXTDuque-Beltrán S., Nicholls-Orejuela R.S., Arévalo-Jamaica A., Guerrero-Lozano R., Montenegro S., James M.A._2002.pdf.txtDuque-Beltrán S., Nicholls-Orejuela R.S., Arévalo-Jamaica A., Guerrero-Lozano R., Montenegro S., James M.A._2002.pdf.txtExtracted texttext/plain23468https://repositorio.unbosque.edu.co/bitstreams/d63d2dac-cf76-40c6-814d-621b7535cdca/downloada83940a80fa7a1b61e800ce4958ae01eMD5520.500.12495/1661oai:repositorio.unbosque.edu.co:20.500.12495/16612024-02-07 05:30:33.684http://creativecommons.org/licenses/by-nc/4.0/Attribution-NonCommercial 4.0 Internationalopen.accesshttps://repositorio.unbosque.edu.coRepositorio Institucional Universidad El Bosquebibliotecas@biteca.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 |