Validation of an indirect in-house ELISA using synthetic peptides to detect antibodies anti-gp90 and gp45 of the equine infectious anaemia virus

Autores
Russi, Romina Cecilia; García, Lucila; Cámara, María Silvia; Soutullo, Adriana Rosa
Año de publicación
2022
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
Background: Equine infectious anaemia (EIA) is controlled by the identification of seropositive animals. The official diagnostic method is the agar gel immunodiffusion (AGID) test, which detects antibodies against a viral core protein (p26). Although AGID is inexpensive and specific, the report of results takes considerable time and the test has low analytical sensitivity. Objective: To validate our in-house indirect ELISAgp90/45, following the World Organization of Animal Health (OIE) criteria. Study design: Test validation. Methods: Synthetic peptides gp90 and gp45 were used as antigens in ELISAgp90/45. Tests used for validation, calibration and linear working operating range, analytical and diagnostic sensitivity and specificity, repeatability and reproducibility were assessed by comparing them with the AGID test and using 1844 equine sera grouped into five different panels. Results: We were able to replace the National References Sera with our Internal Reference Sera. ELISAgp90/45 had acceptable repeatability and reproducibility. Analytical sensitivity of the ELISAgp90/45 was 800 times greater than that of AGID test for positive sera and 400 times greater for weak positive sera. ELISAgp90/45 also showed optimal analytical specificity, since no cross-reactivity was detected with antibodies against other equine viruses. One sample was positive by AGID test and negative by ELISAgp90/45. ELISAgp90/45 was performed using 243 EIA positive and 878 negative equid sera, and showed a diagnostic sensitivity of 99.59% [CI 97.73%-99.99%] and a diagnostic specificity of 90.32% [CI 88.17%-92.19%], compared to AGID test; thus, it was demonstrated to be a robust test. Main limitations: Samples were derived from naturally infected equid populations showing heterogeneous clinical states: therefore, their status was uncertain and some horses were sampled more than once. The AGID test may not be the most useful gold standard. Conclusion: ELISAgp90/45 is a useful tool for the diagnosis of EIAV infection and meets validation requirements established by the OIE.
Fil: Russi, Romina Cecilia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mendoza. Instituto de Medicina y Biología Experimental de Cuyo; Argentina. Universidad Nacional del Litoral. Facultad de Bioquimica y Ciencias Biologicas. Laboratorio de Inmunologia Experimental.; Argentina. Gobierno de la Provincia de Santa Fe. Ministerio de la Producción, Ciencia y Tecnología. Laboratorio de Diagnóstico e Investigaciones Agropecuarias; Argentina
Fil: García, Lucila. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Instituto de Biología Molecular y Celular de Rosario. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Instituto de Biología Molecular y Celular de Rosario; Argentina. Gobierno de la Provincia de Santa Fe. Ministerio de la Producción, Ciencia y Tecnología. Laboratorio de Diagnóstico e Investigaciones Agropecuarias; Argentina
Fil: Cámara, María Silvia. Universidad Nacional del Litoral. Facultad de Bioquímica y Ciencias Biológicas; Argentina
Fil: Soutullo, Adriana Rosa. Gobierno de la Provincia de Santa Fe. Ministerio de la Producción, Ciencia y Tecnología. Laboratorio de Diagnóstico e Investigaciones Agropecuarias; Argentina. Universidad Nacional del Litoral. Facultad de Bioquimica y Ciencias Biologicas. Laboratorio de Inmunologia Experimental.; Argentina
Materia
AGID TEST
ELISA
EQUINE INFECTIOUS ANAEMIA
HORSE
OIE VALIDATION
SYNTHETIC PEPTIDES
Nivel de accesibilidad
acceso abierto
Condiciones de uso
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
Repositorio
CONICET Digital (CONICET)
Institución
Consejo Nacional de Investigaciones Científicas y Técnicas
OAI Identificador
oai:ri.conicet.gov.ar:11336/203439
Acceder
id CONICETDig_f4174f7e857f2efa04d85a679466e4e4
oai_identifier_str oai:ri.conicet.gov.ar:11336/203439
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling Validation of an indirect in-house ELISA using synthetic peptides to detect antibodies anti-gp90 and gp45 of the equine infectious anaemia virusRussi, Romina CeciliaGarcía, LucilaCámara, María SilviaSoutullo, Adriana RosaAGID TESTELISAEQUINE INFECTIOUS ANAEMIAHORSEOIE VALIDATIONSYNTHETIC PEPTIDEShttps://purl.org/becyt/ford/3.3https://purl.org/becyt/ford/3Background: Equine infectious anaemia (EIA) is controlled by the identification of seropositive animals. The official diagnostic method is the agar gel immunodiffusion (AGID) test, which detects antibodies against a viral core protein (p26). Although AGID is inexpensive and specific, the report of results takes considerable time and the test has low analytical sensitivity. Objective: To validate our in-house indirect ELISAgp90/45, following the World Organization of Animal Health (OIE) criteria. Study design: Test validation. Methods: Synthetic peptides gp90 and gp45 were used as antigens in ELISAgp90/45. Tests used for validation, calibration and linear working operating range, analytical and diagnostic sensitivity and specificity, repeatability and reproducibility were assessed by comparing them with the AGID test and using 1844 equine sera grouped into five different panels. Results: We were able to replace the National References Sera with our Internal Reference Sera. ELISAgp90/45 had acceptable repeatability and reproducibility. Analytical sensitivity of the ELISAgp90/45 was 800 times greater than that of AGID test for positive sera and 400 times greater for weak positive sera. ELISAgp90/45 also showed optimal analytical specificity, since no cross-reactivity was detected with antibodies against other equine viruses. One sample was positive by AGID test and negative by ELISAgp90/45. ELISAgp90/45 was performed using 243 EIA positive and 878 negative equid sera, and showed a diagnostic sensitivity of 99.59% [CI 97.73%-99.99%] and a diagnostic specificity of 90.32% [CI 88.17%-92.19%], compared to AGID test; thus, it was demonstrated to be a robust test. Main limitations: Samples were derived from naturally infected equid populations showing heterogeneous clinical states: therefore, their status was uncertain and some horses were sampled more than once. The AGID test may not be the most useful gold standard. Conclusion: ELISAgp90/45 is a useful tool for the diagnosis of EIAV infection and meets validation requirements established by the OIE.Fil: Russi, Romina Cecilia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mendoza. Instituto de Medicina y Biología Experimental de Cuyo; Argentina. Universidad Nacional del Litoral. Facultad de Bioquimica y Ciencias Biologicas. Laboratorio de Inmunologia Experimental.; Argentina. Gobierno de la Provincia de Santa Fe. Ministerio de la Producción, Ciencia y Tecnología. Laboratorio de Diagnóstico e Investigaciones Agropecuarias; ArgentinaFil: García, Lucila. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Instituto de Biología Molecular y Celular de Rosario. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Instituto de Biología Molecular y Celular de Rosario; Argentina. Gobierno de la Provincia de Santa Fe. Ministerio de la Producción, Ciencia y Tecnología. Laboratorio de Diagnóstico e Investigaciones Agropecuarias; ArgentinaFil: Cámara, María Silvia. Universidad Nacional del Litoral. Facultad de Bioquímica y Ciencias Biológicas; ArgentinaFil: Soutullo, Adriana Rosa. Gobierno de la Provincia de Santa Fe. Ministerio de la Producción, Ciencia y Tecnología. Laboratorio de Diagnóstico e Investigaciones Agropecuarias; Argentina. Universidad Nacional del Litoral. Facultad de Bioquimica y Ciencias Biologicas. Laboratorio de Inmunologia Experimental.; ArgentinaEquine Veterinary Journal Ltd2022-01info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/203439Russi, Romina Cecilia; García, Lucila; Cámara, María Silvia; Soutullo, Adriana Rosa; Validation of an indirect in-house ELISA using synthetic peptides to detect antibodies anti-gp90 and gp45 of the equine infectious anaemia virus; Equine Veterinary Journal Ltd; Equine Veterinary Journal; 55; 1; 1-2022; 111-1210425-1644CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/url/https://onlinelibrary.wiley.com/doi/10.1111/evj.13555info:eu-repo/semantics/altIdentifier/doi/10.1111/evj.13555info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-11-12T09:43:18Zoai:ri.conicet.gov.ar:11336/203439instacron:CONICETInstitucionalhttp://ri.conicet.gov.ar/Organismo científico-tecnológicoNo correspondehttp://ri.conicet.gov.ar/oai/requestdasensio@conicet.gov.ar; lcarlino@conicet.gov.arArgentinaNo correspondeNo correspondeNo correspondeopendoar:34982025-11-12 09:43:18.924CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Validation of an indirect in-house ELISA using synthetic peptides to detect antibodies anti-gp90 and gp45 of the equine infectious anaemia virus
title Validation of an indirect in-house ELISA using synthetic peptides to detect antibodies anti-gp90 and gp45 of the equine infectious anaemia virus
spellingShingle Validation of an indirect in-house ELISA using synthetic peptides to detect antibodies anti-gp90 and gp45 of the equine infectious anaemia virus
Russi, Romina Cecilia
AGID TEST
ELISA
EQUINE INFECTIOUS ANAEMIA
HORSE
OIE VALIDATION
SYNTHETIC PEPTIDES
title_short Validation of an indirect in-house ELISA using synthetic peptides to detect antibodies anti-gp90 and gp45 of the equine infectious anaemia virus
title_full Validation of an indirect in-house ELISA using synthetic peptides to detect antibodies anti-gp90 and gp45 of the equine infectious anaemia virus
title_fullStr Validation of an indirect in-house ELISA using synthetic peptides to detect antibodies anti-gp90 and gp45 of the equine infectious anaemia virus
title_full_unstemmed Validation of an indirect in-house ELISA using synthetic peptides to detect antibodies anti-gp90 and gp45 of the equine infectious anaemia virus
title_sort Validation of an indirect in-house ELISA using synthetic peptides to detect antibodies anti-gp90 and gp45 of the equine infectious anaemia virus
dc.creator.none.fl_str_mv Russi, Romina Cecilia
García, Lucila
Cámara, María Silvia
Soutullo, Adriana Rosa
author Russi, Romina Cecilia
author_facet Russi, Romina Cecilia
García, Lucila
Cámara, María Silvia
Soutullo, Adriana Rosa
author_role author
author2 García, Lucila
Cámara, María Silvia
Soutullo, Adriana Rosa
author2_role author
author
author
dc.subject.none.fl_str_mv AGID TEST
ELISA
EQUINE INFECTIOUS ANAEMIA
HORSE
OIE VALIDATION
SYNTHETIC PEPTIDES
topic AGID TEST
ELISA
EQUINE INFECTIOUS ANAEMIA
HORSE
OIE VALIDATION
SYNTHETIC PEPTIDES
purl_subject.fl_str_mv https://purl.org/becyt/ford/3.3
https://purl.org/becyt/ford/3
dc.description.none.fl_txt_mv Background: Equine infectious anaemia (EIA) is controlled by the identification of seropositive animals. The official diagnostic method is the agar gel immunodiffusion (AGID) test, which detects antibodies against a viral core protein (p26). Although AGID is inexpensive and specific, the report of results takes considerable time and the test has low analytical sensitivity. Objective: To validate our in-house indirect ELISAgp90/45, following the World Organization of Animal Health (OIE) criteria. Study design: Test validation. Methods: Synthetic peptides gp90 and gp45 were used as antigens in ELISAgp90/45. Tests used for validation, calibration and linear working operating range, analytical and diagnostic sensitivity and specificity, repeatability and reproducibility were assessed by comparing them with the AGID test and using 1844 equine sera grouped into five different panels. Results: We were able to replace the National References Sera with our Internal Reference Sera. ELISAgp90/45 had acceptable repeatability and reproducibility. Analytical sensitivity of the ELISAgp90/45 was 800 times greater than that of AGID test for positive sera and 400 times greater for weak positive sera. ELISAgp90/45 also showed optimal analytical specificity, since no cross-reactivity was detected with antibodies against other equine viruses. One sample was positive by AGID test and negative by ELISAgp90/45. ELISAgp90/45 was performed using 243 EIA positive and 878 negative equid sera, and showed a diagnostic sensitivity of 99.59% [CI 97.73%-99.99%] and a diagnostic specificity of 90.32% [CI 88.17%-92.19%], compared to AGID test; thus, it was demonstrated to be a robust test. Main limitations: Samples were derived from naturally infected equid populations showing heterogeneous clinical states: therefore, their status was uncertain and some horses were sampled more than once. The AGID test may not be the most useful gold standard. Conclusion: ELISAgp90/45 is a useful tool for the diagnosis of EIAV infection and meets validation requirements established by the OIE.
Fil: Russi, Romina Cecilia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mendoza. Instituto de Medicina y Biología Experimental de Cuyo; Argentina. Universidad Nacional del Litoral. Facultad de Bioquimica y Ciencias Biologicas. Laboratorio de Inmunologia Experimental.; Argentina. Gobierno de la Provincia de Santa Fe. Ministerio de la Producción, Ciencia y Tecnología. Laboratorio de Diagnóstico e Investigaciones Agropecuarias; Argentina
Fil: García, Lucila. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Instituto de Biología Molecular y Celular de Rosario. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Instituto de Biología Molecular y Celular de Rosario; Argentina. Gobierno de la Provincia de Santa Fe. Ministerio de la Producción, Ciencia y Tecnología. Laboratorio de Diagnóstico e Investigaciones Agropecuarias; Argentina
Fil: Cámara, María Silvia. Universidad Nacional del Litoral. Facultad de Bioquímica y Ciencias Biológicas; Argentina
Fil: Soutullo, Adriana Rosa. Gobierno de la Provincia de Santa Fe. Ministerio de la Producción, Ciencia y Tecnología. Laboratorio de Diagnóstico e Investigaciones Agropecuarias; Argentina. Universidad Nacional del Litoral. Facultad de Bioquimica y Ciencias Biologicas. Laboratorio de Inmunologia Experimental.; Argentina
description Background: Equine infectious anaemia (EIA) is controlled by the identification of seropositive animals. The official diagnostic method is the agar gel immunodiffusion (AGID) test, which detects antibodies against a viral core protein (p26). Although AGID is inexpensive and specific, the report of results takes considerable time and the test has low analytical sensitivity. Objective: To validate our in-house indirect ELISAgp90/45, following the World Organization of Animal Health (OIE) criteria. Study design: Test validation. Methods: Synthetic peptides gp90 and gp45 were used as antigens in ELISAgp90/45. Tests used for validation, calibration and linear working operating range, analytical and diagnostic sensitivity and specificity, repeatability and reproducibility were assessed by comparing them with the AGID test and using 1844 equine sera grouped into five different panels. Results: We were able to replace the National References Sera with our Internal Reference Sera. ELISAgp90/45 had acceptable repeatability and reproducibility. Analytical sensitivity of the ELISAgp90/45 was 800 times greater than that of AGID test for positive sera and 400 times greater for weak positive sera. ELISAgp90/45 also showed optimal analytical specificity, since no cross-reactivity was detected with antibodies against other equine viruses. One sample was positive by AGID test and negative by ELISAgp90/45. ELISAgp90/45 was performed using 243 EIA positive and 878 negative equid sera, and showed a diagnostic sensitivity of 99.59% [CI 97.73%-99.99%] and a diagnostic specificity of 90.32% [CI 88.17%-92.19%], compared to AGID test; thus, it was demonstrated to be a robust test. Main limitations: Samples were derived from naturally infected equid populations showing heterogeneous clinical states: therefore, their status was uncertain and some horses were sampled more than once. The AGID test may not be the most useful gold standard. Conclusion: ELISAgp90/45 is a useful tool for the diagnosis of EIAV infection and meets validation requirements established by the OIE.
publishDate 2022
dc.date.none.fl_str_mv 2022-01
dc.type.none.fl_str_mv info:eu-repo/semantics/article
info:eu-repo/semantics/publishedVersion
http://purl.org/coar/resource_type/c_6501
info:ar-repo/semantics/articulo
format article
status_str publishedVersion
dc.identifier.none.fl_str_mv http://hdl.handle.net/11336/203439
Russi, Romina Cecilia; García, Lucila; Cámara, María Silvia; Soutullo, Adriana Rosa; Validation of an indirect in-house ELISA using synthetic peptides to detect antibodies anti-gp90 and gp45 of the equine infectious anaemia virus; Equine Veterinary Journal Ltd; Equine Veterinary Journal; 55; 1; 1-2022; 111-121
0425-1644
CONICET Digital
CONICET
url http://hdl.handle.net/11336/203439
identifier_str_mv Russi, Romina Cecilia; García, Lucila; Cámara, María Silvia; Soutullo, Adriana Rosa; Validation of an indirect in-house ELISA using synthetic peptides to detect antibodies anti-gp90 and gp45 of the equine infectious anaemia virus; Equine Veterinary Journal Ltd; Equine Veterinary Journal; 55; 1; 1-2022; 111-121
0425-1644
CONICET Digital
CONICET
dc.language.none.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv info:eu-repo/semantics/altIdentifier/url/https://onlinelibrary.wiley.com/doi/10.1111/evj.13555
info:eu-repo/semantics/altIdentifier/doi/10.1111/evj.13555
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
eu_rights_str_mv openAccess
rights_invalid_str_mv https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
dc.format.none.fl_str_mv application/pdf
application/pdf
application/pdf
dc.publisher.none.fl_str_mv Equine Veterinary Journal Ltd
publisher.none.fl_str_mv Equine Veterinary Journal Ltd
dc.source.none.fl_str_mv reponame:CONICET Digital (CONICET)
instname:Consejo Nacional de Investigaciones Científicas y Técnicas
reponame_str CONICET Digital (CONICET)
collection CONICET Digital (CONICET)
instname_str Consejo Nacional de Investigaciones Científicas y Técnicas
repository.name.fl_str_mv CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicas
repository.mail.fl_str_mv dasensio@conicet.gov.ar; lcarlino@conicet.gov.ar
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score 13.24909

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