ELISA technique standardization for strongyloidiasis diagnosis

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Objective: To standardize ELISA technique for human Strongyloides stercoralis infection diagnosis. Material and methods: A crude antigen was prepared using filariform larvae obtained from positive stool samples cultured with charcoal. Harvested larvae were crushed by sonication and washed by centrif...

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Detalles Bibliográficos
Autores: Huapaya, Pedro, Espinoza, Yrma, Huiza, Alina, Sevilla, Carlos
Formato: artículo
Fecha de Publicación:2002
Institución:Universidad Nacional Mayor de San Marcos
Repositorio:Revistas - Universidad Nacional Mayor de San Marcos
Lenguaje:español
OAI Identifier:oai:ojs.csi.unmsm:article/1496
Enlace del recurso:https://revistasinvestigacion.unmsm.edu.pe/index.php/anales/article/view/1496
Nivel de acceso:acceso abierto
Materia:Strongyloides stercoralis
estrongiloidiosis
ELISA
serología
diagnóstico de laboratorio
strongyloidiasis
serology
diagnosis laboratory.
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spelling ELISA technique standardization for strongyloidiasis diagnosisEstandarización de la técnica de ELISA para diagnóstico de estrongiloidiosisHuapaya, PedroEspinoza, YrmaHuiza, AlinaSevilla, CarlosStrongyloides stercoralisestrongiloidiosisELISAserologíadiagnóstico de laboratorioStrongyloides stercoralisstrongyloidiasisELISAserologydiagnosis laboratory.Objective: To standardize ELISA technique for human Strongyloides stercoralis infection diagnosis. Material and methods: A crude antigen was prepared using filariform larvae obtained from positive stool samples cultured with charcoal. Harvested larvae were crushed by sonication and washed by centrifugation in order to obtain protein extracts to be used as antigen. Final protein concentration was 600 µg/mL. Several kinds of ELISA plates were tested and antigen concentration, sera dilution, conjugate dilution and cut off were determined to identify infection. Sera from patients with both hyperinfection syndrome and intestinal infection demonstrated by parasitological examination were positive controls and sera from people living in non-endemic areas with no infection demonstrated by parasitological examination were negative controls. Results: Best values were 5 µg/mL for antigen, 1/64 for sera, 1/1000 for conjugate; optical density values for positive samples were 1,2746 (1,1065 – 1,4206, DS = 0,3284) and for negative samples 0,4457 (0,3324 – 0,5538, DS = 0,2230). Twenty sera samples from positive subjects and one hundred from negative subjects were examined, obtaining 90% sensitivity and 88% specificity. Conclusion: The results show this technique could be useful as strongyloidiasis screening test in population studies.Objetivo: Estandarizar la técnica de ELISA para diagnóstico de la infección humana por el parásito Strongyloides stercoralis. Material y métodos: Se preparó antígeno crudo usando larvas filariformes obtenidas de muestras de heces positivas cultivadas con carbón vegetal. Las larvas fueron trituradas mediante sonicación y lavadas por centrifugación para obtener extractos de proteínas para usarlos como antígeno. La concentración proteica final fue de 600 µg/mL. Se probó varios tipos de placas de ELISA y se determinó las concentraciones de antígeno, sueros, conjugado y puntos de corte, para permitir la diferenciación de la infección. Los controles positivos fueron sueros de pacientes con síndrome de hiperinfección e infección intestinal, comprobados mediante exámenes parasitológicos de heces; y los controles negativos fueron sueros de personas provenientes de zonas no endémicas y con comprobación mediante exámenes parasitológicos de la ausencia del parásito. Resultados: Los valores óptimos fueron 5 µg/mL para el antígeno, 1/64 para el suero, 1/1000 para el conjugado; los valores de densidad óptica para las muestras positivas fueron en promedio 1,2746 (1,1065 – 1,4206, DS = 0,3284) y de las muestras negativas 0,4457 (0,3324 – 0,5538, DS = 0,2230). Se examinó 20 muestras de suero de sujetos positivos y 100 de sujetos negativos, obteniéndose sensibilidad de 90% y especificidad de 88%. Conclusión: Los resultados muestran que esta técnica puede constituirse en una prueba de tamizaje de estrongiloidiosis en estudios de población.Universidad Nacional Mayor de San Marcos, Facultad de Medicina Humana2002-09-16info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionapplication/pdfhttps://revistasinvestigacion.unmsm.edu.pe/index.php/anales/article/view/149610.15381/anales.v63i3.1496Anales de la Facultad de Medicina; Vol. 63 No. 3 (2002); 179-184Anales de la Facultad de Medicina; Vol. 63 Núm. 3 (2002); 179-1841609-94191025-5583reponame:Revistas - Universidad Nacional Mayor de San Marcosinstname:Universidad Nacional Mayor de San Marcosinstacron:UNMSMspahttps://revistasinvestigacion.unmsm.edu.pe/index.php/anales/article/view/1496/12196Derechos de autor 2002 Pedro huapaya, Yrma Espinoza, Alina Huiza, Carlos Sevillahttps://creativecommons.org/licenses/by-nc-sa/4.0info:eu-repo/semantics/openAccessoai:ojs.csi.unmsm:article/14962020-04-14T19:05:49Z
dc.title.none.fl_str_mv ELISA technique standardization for strongyloidiasis diagnosis
Estandarización de la técnica de ELISA para diagnóstico de estrongiloidiosis
title ELISA technique standardization for strongyloidiasis diagnosis
spellingShingle ELISA technique standardization for strongyloidiasis diagnosis
Huapaya, Pedro
Strongyloides stercoralis
estrongiloidiosis
ELISA
serología
diagnóstico de laboratorio
Strongyloides stercoralis
strongyloidiasis
ELISA
serology
diagnosis laboratory.
title_short ELISA technique standardization for strongyloidiasis diagnosis
title_full ELISA technique standardization for strongyloidiasis diagnosis
title_fullStr ELISA technique standardization for strongyloidiasis diagnosis
title_full_unstemmed ELISA technique standardization for strongyloidiasis diagnosis
title_sort ELISA technique standardization for strongyloidiasis diagnosis
dc.creator.none.fl_str_mv Huapaya, Pedro
Espinoza, Yrma
Huiza, Alina
Sevilla, Carlos
author Huapaya, Pedro
author_facet Huapaya, Pedro
Espinoza, Yrma
Huiza, Alina
Sevilla, Carlos
author_role author
author2 Espinoza, Yrma
Huiza, Alina
Sevilla, Carlos
author2_role author
author
author
dc.subject.none.fl_str_mv Strongyloides stercoralis
estrongiloidiosis
ELISA
serología
diagnóstico de laboratorio
Strongyloides stercoralis
strongyloidiasis
ELISA
serology
diagnosis laboratory.
topic Strongyloides stercoralis
estrongiloidiosis
ELISA
serología
diagnóstico de laboratorio
Strongyloides stercoralis
strongyloidiasis
ELISA
serology
diagnosis laboratory.
description Objective: To standardize ELISA technique for human Strongyloides stercoralis infection diagnosis. Material and methods: A crude antigen was prepared using filariform larvae obtained from positive stool samples cultured with charcoal. Harvested larvae were crushed by sonication and washed by centrifugation in order to obtain protein extracts to be used as antigen. Final protein concentration was 600 µg/mL. Several kinds of ELISA plates were tested and antigen concentration, sera dilution, conjugate dilution and cut off were determined to identify infection. Sera from patients with both hyperinfection syndrome and intestinal infection demonstrated by parasitological examination were positive controls and sera from people living in non-endemic areas with no infection demonstrated by parasitological examination were negative controls. Results: Best values were 5 µg/mL for antigen, 1/64 for sera, 1/1000 for conjugate; optical density values for positive samples were 1,2746 (1,1065 – 1,4206, DS = 0,3284) and for negative samples 0,4457 (0,3324 – 0,5538, DS = 0,2230). Twenty sera samples from positive subjects and one hundred from negative subjects were examined, obtaining 90% sensitivity and 88% specificity. Conclusion: The results show this technique could be useful as strongyloidiasis screening test in population studies.
publishDate 2002
dc.date.none.fl_str_mv 2002-09-16
dc.type.none.fl_str_mv info:eu-repo/semantics/article
info:eu-repo/semantics/publishedVersion
format article
status_str publishedVersion
dc.identifier.none.fl_str_mv https://revistasinvestigacion.unmsm.edu.pe/index.php/anales/article/view/1496
10.15381/anales.v63i3.1496
url https://revistasinvestigacion.unmsm.edu.pe/index.php/anales/article/view/1496
identifier_str_mv 10.15381/anales.v63i3.1496
dc.language.none.fl_str_mv spa
language spa
dc.relation.none.fl_str_mv https://revistasinvestigacion.unmsm.edu.pe/index.php/anales/article/view/1496/12196
dc.rights.none.fl_str_mv Derechos de autor 2002 Pedro huapaya, Yrma Espinoza, Alina Huiza, Carlos Sevilla
https://creativecommons.org/licenses/by-nc-sa/4.0
info:eu-repo/semantics/openAccess
rights_invalid_str_mv Derechos de autor 2002 Pedro huapaya, Yrma Espinoza, Alina Huiza, Carlos Sevilla
https://creativecommons.org/licenses/by-nc-sa/4.0
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv application/pdf
dc.publisher.none.fl_str_mv Universidad Nacional Mayor de San Marcos, Facultad de Medicina Humana
publisher.none.fl_str_mv Universidad Nacional Mayor de San Marcos, Facultad de Medicina Humana
dc.source.none.fl_str_mv Anales de la Facultad de Medicina; Vol. 63 No. 3 (2002); 179-184
Anales de la Facultad de Medicina; Vol. 63 Núm. 3 (2002); 179-184
1609-9419
1025-5583
reponame:Revistas - Universidad Nacional Mayor de San Marcos
instname:Universidad Nacional Mayor de San Marcos
instacron:UNMSM
instname_str Universidad Nacional Mayor de San Marcos
instacron_str UNMSM
institution UNMSM
reponame_str Revistas - Universidad Nacional Mayor de San Marcos
collection Revistas - Universidad Nacional Mayor de San Marcos
repository.name.fl_str_mv
repository.mail.fl_str_mv
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score 13.904009
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