Development and validation of an Enzyme-Linked Immunosorbent Assay for the detection of antibodies against Trichinella spp. in domestic pigs in Argentina

Autores
Riva, Eliana; Steffan, Pedro Eduardo; Amanto, Andres Fabian; Davila, Agustin; Bernat, Gisele; Fernández, Alicia Silvina; Fuentes, Mariana Elisabet; Estein, Silvia Marcela
Año de publicación
2021
Idioma
inglés
Tipo de recurso
documento de conferencia
Estado
versión publicada
Descripción
Trichinellosis is endemic in Argentina, with human outbreaks having mainly domestic pigs as focalised sources of infection. Enzyme-Linked Immunosorbent Assay (ELISA), based on excretory/secretory T. spiralis muscle larvae antigens (E/S ML) is the most recommended technique for epidemiological surveillance to detect anti-Trichinella antibodies in domestic pigs. Available international ELISA kits are expensive and need to be validated in each country. Thus, the aim of the present study was to evaluate the performance of an in-house indirect ELISA based on E/S ML antigen. The artificial digestion (AD) was used as reference technique. A total of 343 swine serum samples were used to define sensitivity, specificity, accuracy and repeatability of the ELISA. Sera tested were from 91 outdoor pigs associated with local outbreaks, 12 of them diagnosed as positive by AD, and 242 serum samples from intensive-rearing farms with strict hygienic conditions and negative results by AD. Sera from 2 pigs experimentally infected with 10,000 T. spiralis ML (bled at five different days post infection) were also included. E/S ML antigen was diluted at 5 μg/ml in coating buffer, samples were diluted 1/100 in wash buffer and peroxidase conjugated rabbit anti-pig IgG was used at 1/2500 dilution (Sigma). Reaction was developed using ortho-phenylene-diamine diluted in a citric acid buffer 0.07% H2O2. After 15 minutes, reaction was stopped with H2SO4 2.5N. The cut-off point was calculated based on the OD values of 5 negative controls (NC) using the formula: Mean NC ± 3 SD. The diagnostic sensitivity and specificity of the ELISA were 95.5% and 98.4%, respectively. The accuracy was 98.2%. The inter-assay repeatability (4 plates) calculated for 1 positive and 4 negative controls was under 20%. The performance of this ELISA suggests that it could be used as a reliable and practical tool for seroepidemiological studies in Argentina.
Fil: Riva, Eliana. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tandil. Centro de Investigación Veterinaria de Tandil. Universidad Nacional del Centro de la Provincia de Buenos Aires. Centro de Investigación Veterinaria de Tandil. Provincia de Buenos Aires. Gobernación. Comision de Investigaciones Científicas. Centro de Investigación Veterinaria de Tandil; Argentina
Fil: Steffan, Pedro Eduardo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tandil. Centro de Investigación Veterinaria de Tandil. Universidad Nacional del Centro de la Provincia de Buenos Aires. Centro de Investigación Veterinaria de Tandil. Provincia de Buenos Aires. Gobernación. Comision de Investigaciones Científicas. Centro de Investigación Veterinaria de Tandil; Argentina
Fil: Amanto, Andres Fabian. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tandil. Centro de Investigación Veterinaria de Tandil. Universidad Nacional del Centro de la Provincia de Buenos Aires. Centro de Investigación Veterinaria de Tandil. Provincia de Buenos Aires. Gobernación. Comision de Investigaciones Científicas. Centro de Investigación Veterinaria de Tandil; Argentina
Fil: Davila, Agustin. Universidad Nacional del Centro de la Provincia de Buenos Aires. Facultad de Ciencias Veterinarias; Argentina
Fil: Bernat, Gisele. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tandil. Centro de Investigación Veterinaria de Tandil. Universidad Nacional del Centro de la Provincia de Buenos Aires. Centro de Investigación Veterinaria de Tandil. Provincia de Buenos Aires. Gobernación. Comision de Investigaciones Científicas. Centro de Investigación Veterinaria de Tandil; Argentina
Fil: Fernández, Alicia Silvina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tandil. Centro de Investigación Veterinaria de Tandil. Universidad Nacional del Centro de la Provincia de Buenos Aires. Centro de Investigación Veterinaria de Tandil. Provincia de Buenos Aires. Gobernación. Comision de Investigaciones Científicas. Centro de Investigación Veterinaria de Tandil; Argentina
Fil: Fuentes, Mariana Elisabet. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tandil. Centro de Investigación Veterinaria de Tandil. Universidad Nacional del Centro de la Provincia de Buenos Aires. Centro de Investigación Veterinaria de Tandil. Provincia de Buenos Aires. Gobernación. Comision de Investigaciones Científicas. Centro de Investigación Veterinaria de Tandil; Argentina
Fil: Estein, Silvia Marcela. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tandil. Centro de Investigación Veterinaria de Tandil. Universidad Nacional del Centro de la Provincia de Buenos Aires. Centro de Investigación Veterinaria de Tandil. Provincia de Buenos Aires. Gobernación. Comision de Investigaciones Científicas. Centro de Investigación Veterinaria de Tandil; Argentina
28th International Conference of the World Association for the Advancement of Veterinary Parasitology
Dublin
Reino Unido
University College of Dublin
Materia
TRICHINELLOSIS
DIAGNOSTICO
ELISA
CERDOS
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:11336/156172
Acceder
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spelling Development and validation of an Enzyme-Linked Immunosorbent Assay for the detection of antibodies against Trichinella spp. in domestic pigs in ArgentinaRiva, ElianaSteffan, Pedro EduardoAmanto, Andres FabianDavila, AgustinBernat, GiseleFernández, Alicia SilvinaFuentes, Mariana ElisabetEstein, Silvia MarcelaTRICHINELLOSISDIAGNOSTICOELISACERDOShttps://purl.org/becyt/ford/4.3https://purl.org/becyt/ford/4Trichinellosis is endemic in Argentina, with human outbreaks having mainly domestic pigs as focalised sources of infection. Enzyme-Linked Immunosorbent Assay (ELISA), based on excretory/secretory T. spiralis muscle larvae antigens (E/S ML) is the most recommended technique for epidemiological surveillance to detect anti-Trichinella antibodies in domestic pigs. Available international ELISA kits are expensive and need to be validated in each country. Thus, the aim of the present study was to evaluate the performance of an in-house indirect ELISA based on E/S ML antigen. The artificial digestion (AD) was used as reference technique. A total of 343 swine serum samples were used to define sensitivity, specificity, accuracy and repeatability of the ELISA. Sera tested were from 91 outdoor pigs associated with local outbreaks, 12 of them diagnosed as positive by AD, and 242 serum samples from intensive-rearing farms with strict hygienic conditions and negative results by AD. Sera from 2 pigs experimentally infected with 10,000 T. spiralis ML (bled at five different days post infection) were also included. E/S ML antigen was diluted at 5 μg/ml in coating buffer, samples were diluted 1/100 in wash buffer and peroxidase conjugated rabbit anti-pig IgG was used at 1/2500 dilution (Sigma). Reaction was developed using ortho-phenylene-diamine diluted in a citric acid buffer 0.07% H2O2. After 15 minutes, reaction was stopped with H2SO4 2.5N. The cut-off point was calculated based on the OD values of 5 negative controls (NC) using the formula: Mean NC ± 3 SD. The diagnostic sensitivity and specificity of the ELISA were 95.5% and 98.4%, respectively. The accuracy was 98.2%. The inter-assay repeatability (4 plates) calculated for 1 positive and 4 negative controls was under 20%. The performance of this ELISA suggests that it could be used as a reliable and practical tool for seroepidemiological studies in Argentina.Fil: Riva, Eliana. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tandil. Centro de Investigación Veterinaria de Tandil. Universidad Nacional del Centro de la Provincia de Buenos Aires. Centro de Investigación Veterinaria de Tandil. Provincia de Buenos Aires. Gobernación. Comision de Investigaciones Científicas. Centro de Investigación Veterinaria de Tandil; ArgentinaFil: Steffan, Pedro Eduardo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tandil. Centro de Investigación Veterinaria de Tandil. Universidad Nacional del Centro de la Provincia de Buenos Aires. Centro de Investigación Veterinaria de Tandil. Provincia de Buenos Aires. Gobernación. Comision de Investigaciones Científicas. Centro de Investigación Veterinaria de Tandil; ArgentinaFil: Amanto, Andres Fabian. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tandil. Centro de Investigación Veterinaria de Tandil. Universidad Nacional del Centro de la Provincia de Buenos Aires. Centro de Investigación Veterinaria de Tandil. Provincia de Buenos Aires. Gobernación. Comision de Investigaciones Científicas. Centro de Investigación Veterinaria de Tandil; ArgentinaFil: Davila, Agustin. Universidad Nacional del Centro de la Provincia de Buenos Aires. Facultad de Ciencias Veterinarias; ArgentinaFil: Bernat, Gisele. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tandil. Centro de Investigación Veterinaria de Tandil. Universidad Nacional del Centro de la Provincia de Buenos Aires. Centro de Investigación Veterinaria de Tandil. Provincia de Buenos Aires. Gobernación. Comision de Investigaciones Científicas. Centro de Investigación Veterinaria de Tandil; ArgentinaFil: Fernández, Alicia Silvina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tandil. Centro de Investigación Veterinaria de Tandil. Universidad Nacional del Centro de la Provincia de Buenos Aires. Centro de Investigación Veterinaria de Tandil. Provincia de Buenos Aires. Gobernación. Comision de Investigaciones Científicas. Centro de Investigación Veterinaria de Tandil; ArgentinaFil: Fuentes, Mariana Elisabet. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tandil. Centro de Investigación Veterinaria de Tandil. Universidad Nacional del Centro de la Provincia de Buenos Aires. Centro de Investigación Veterinaria de Tandil. Provincia de Buenos Aires. Gobernación. Comision de Investigaciones Científicas. Centro de Investigación Veterinaria de Tandil; ArgentinaFil: Estein, Silvia Marcela. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tandil. Centro de Investigación Veterinaria de Tandil. Universidad Nacional del Centro de la Provincia de Buenos Aires. Centro de Investigación Veterinaria de Tandil. Provincia de Buenos Aires. Gobernación. Comision de Investigaciones Científicas. Centro de Investigación Veterinaria de Tandil; Argentina28th International Conference of the World Association for the Advancement of Veterinary ParasitologyDublinReino UnidoUniversity College of DublinWorld Association for the Advancement of Veterinary Parasitology2021info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/conferenceObjectVirtualBookhttp://purl.org/coar/resource_type/c_5794info:ar-repo/semantics/documentoDeConferenciaapplication/pdfapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/156172Development and validation of an Enzyme-Linked Immunosorbent Assay for the detection of antibodies against Trichinella spp. in domestic pigs in Argentina; 28th International Conference of the World Association for the Advancement of Veterinary Parasitology; Dublin; Reino Unido; 2021; 452-452CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/url/http://www.waavp2021.cominfo:eu-repo/semantics/altIdentifier/url/http://www.waavp2021.com/wp-content/uploads/2021/08/WAAVP-2021-Abstract-book-22.07.21-FINAL.pdfInternacionalinfo:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-09-29T10:14:52Zoai:ri.conicet.gov.ar:11336/156172instacron: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:34982025-09-29 10:14:52.333CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Development and validation of an Enzyme-Linked Immunosorbent Assay for the detection of antibodies against Trichinella spp. in domestic pigs in Argentina
title Development and validation of an Enzyme-Linked Immunosorbent Assay for the detection of antibodies against Trichinella spp. in domestic pigs in Argentina
spellingShingle Development and validation of an Enzyme-Linked Immunosorbent Assay for the detection of antibodies against Trichinella spp. in domestic pigs in Argentina
Riva, Eliana
TRICHINELLOSIS
DIAGNOSTICO
ELISA
CERDOS
title_short Development and validation of an Enzyme-Linked Immunosorbent Assay for the detection of antibodies against Trichinella spp. in domestic pigs in Argentina
title_full Development and validation of an Enzyme-Linked Immunosorbent Assay for the detection of antibodies against Trichinella spp. in domestic pigs in Argentina
title_fullStr Development and validation of an Enzyme-Linked Immunosorbent Assay for the detection of antibodies against Trichinella spp. in domestic pigs in Argentina
title_full_unstemmed Development and validation of an Enzyme-Linked Immunosorbent Assay for the detection of antibodies against Trichinella spp. in domestic pigs in Argentina
title_sort Development and validation of an Enzyme-Linked Immunosorbent Assay for the detection of antibodies against Trichinella spp. in domestic pigs in Argentina
dc.creator.none.fl_str_mv Riva, Eliana
Steffan, Pedro Eduardo
Amanto, Andres Fabian
Davila, Agustin
Bernat, Gisele
Fernández, Alicia Silvina
Fuentes, Mariana Elisabet
Estein, Silvia Marcela
author Riva, Eliana
author_facet Riva, Eliana
Steffan, Pedro Eduardo
Amanto, Andres Fabian
Davila, Agustin
Bernat, Gisele
Fernández, Alicia Silvina
Fuentes, Mariana Elisabet
Estein, Silvia Marcela
author_role author
author2 Steffan, Pedro Eduardo
Amanto, Andres Fabian
Davila, Agustin
Bernat, Gisele
Fernández, Alicia Silvina
Fuentes, Mariana Elisabet
Estein, Silvia Marcela
author2_role author
author
author
author
author
author
author
dc.subject.none.fl_str_mv TRICHINELLOSIS
DIAGNOSTICO
ELISA
CERDOS
topic TRICHINELLOSIS
DIAGNOSTICO
ELISA
CERDOS
purl_subject.fl_str_mv https://purl.org/becyt/ford/4.3
https://purl.org/becyt/ford/4
dc.description.none.fl_txt_mv Trichinellosis is endemic in Argentina, with human outbreaks having mainly domestic pigs as focalised sources of infection. Enzyme-Linked Immunosorbent Assay (ELISA), based on excretory/secretory T. spiralis muscle larvae antigens (E/S ML) is the most recommended technique for epidemiological surveillance to detect anti-Trichinella antibodies in domestic pigs. Available international ELISA kits are expensive and need to be validated in each country. Thus, the aim of the present study was to evaluate the performance of an in-house indirect ELISA based on E/S ML antigen. The artificial digestion (AD) was used as reference technique. A total of 343 swine serum samples were used to define sensitivity, specificity, accuracy and repeatability of the ELISA. Sera tested were from 91 outdoor pigs associated with local outbreaks, 12 of them diagnosed as positive by AD, and 242 serum samples from intensive-rearing farms with strict hygienic conditions and negative results by AD. Sera from 2 pigs experimentally infected with 10,000 T. spiralis ML (bled at five different days post infection) were also included. E/S ML antigen was diluted at 5 μg/ml in coating buffer, samples were diluted 1/100 in wash buffer and peroxidase conjugated rabbit anti-pig IgG was used at 1/2500 dilution (Sigma). Reaction was developed using ortho-phenylene-diamine diluted in a citric acid buffer 0.07% H2O2. After 15 minutes, reaction was stopped with H2SO4 2.5N. The cut-off point was calculated based on the OD values of 5 negative controls (NC) using the formula: Mean NC ± 3 SD. The diagnostic sensitivity and specificity of the ELISA were 95.5% and 98.4%, respectively. The accuracy was 98.2%. The inter-assay repeatability (4 plates) calculated for 1 positive and 4 negative controls was under 20%. The performance of this ELISA suggests that it could be used as a reliable and practical tool for seroepidemiological studies in Argentina.
Fil: Riva, Eliana. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tandil. Centro de Investigación Veterinaria de Tandil. Universidad Nacional del Centro de la Provincia de Buenos Aires. Centro de Investigación Veterinaria de Tandil. Provincia de Buenos Aires. Gobernación. Comision de Investigaciones Científicas. Centro de Investigación Veterinaria de Tandil; Argentina
Fil: Steffan, Pedro Eduardo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tandil. Centro de Investigación Veterinaria de Tandil. Universidad Nacional del Centro de la Provincia de Buenos Aires. Centro de Investigación Veterinaria de Tandil. Provincia de Buenos Aires. Gobernación. Comision de Investigaciones Científicas. Centro de Investigación Veterinaria de Tandil; Argentina
Fil: Amanto, Andres Fabian. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tandil. Centro de Investigación Veterinaria de Tandil. Universidad Nacional del Centro de la Provincia de Buenos Aires. Centro de Investigación Veterinaria de Tandil. Provincia de Buenos Aires. Gobernación. Comision de Investigaciones Científicas. Centro de Investigación Veterinaria de Tandil; Argentina
Fil: Davila, Agustin. Universidad Nacional del Centro de la Provincia de Buenos Aires. Facultad de Ciencias Veterinarias; Argentina
Fil: Bernat, Gisele. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tandil. Centro de Investigación Veterinaria de Tandil. Universidad Nacional del Centro de la Provincia de Buenos Aires. Centro de Investigación Veterinaria de Tandil. Provincia de Buenos Aires. Gobernación. Comision de Investigaciones Científicas. Centro de Investigación Veterinaria de Tandil; Argentina
Fil: Fernández, Alicia Silvina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tandil. Centro de Investigación Veterinaria de Tandil. Universidad Nacional del Centro de la Provincia de Buenos Aires. Centro de Investigación Veterinaria de Tandil. Provincia de Buenos Aires. Gobernación. Comision de Investigaciones Científicas. Centro de Investigación Veterinaria de Tandil; Argentina
Fil: Fuentes, Mariana Elisabet. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tandil. Centro de Investigación Veterinaria de Tandil. Universidad Nacional del Centro de la Provincia de Buenos Aires. Centro de Investigación Veterinaria de Tandil. Provincia de Buenos Aires. Gobernación. Comision de Investigaciones Científicas. Centro de Investigación Veterinaria de Tandil; Argentina
Fil: Estein, Silvia Marcela. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tandil. Centro de Investigación Veterinaria de Tandil. Universidad Nacional del Centro de la Provincia de Buenos Aires. Centro de Investigación Veterinaria de Tandil. Provincia de Buenos Aires. Gobernación. Comision de Investigaciones Científicas. Centro de Investigación Veterinaria de Tandil; Argentina
28th International Conference of the World Association for the Advancement of Veterinary Parasitology
Dublin
Reino Unido
University College of Dublin
description Trichinellosis is endemic in Argentina, with human outbreaks having mainly domestic pigs as focalised sources of infection. Enzyme-Linked Immunosorbent Assay (ELISA), based on excretory/secretory T. spiralis muscle larvae antigens (E/S ML) is the most recommended technique for epidemiological surveillance to detect anti-Trichinella antibodies in domestic pigs. Available international ELISA kits are expensive and need to be validated in each country. Thus, the aim of the present study was to evaluate the performance of an in-house indirect ELISA based on E/S ML antigen. The artificial digestion (AD) was used as reference technique. A total of 343 swine serum samples were used to define sensitivity, specificity, accuracy and repeatability of the ELISA. Sera tested were from 91 outdoor pigs associated with local outbreaks, 12 of them diagnosed as positive by AD, and 242 serum samples from intensive-rearing farms with strict hygienic conditions and negative results by AD. Sera from 2 pigs experimentally infected with 10,000 T. spiralis ML (bled at five different days post infection) were also included. E/S ML antigen was diluted at 5 μg/ml in coating buffer, samples were diluted 1/100 in wash buffer and peroxidase conjugated rabbit anti-pig IgG was used at 1/2500 dilution (Sigma). Reaction was developed using ortho-phenylene-diamine diluted in a citric acid buffer 0.07% H2O2. After 15 minutes, reaction was stopped with H2SO4 2.5N. The cut-off point was calculated based on the OD values of 5 negative controls (NC) using the formula: Mean NC ± 3 SD. The diagnostic sensitivity and specificity of the ELISA were 95.5% and 98.4%, respectively. The accuracy was 98.2%. The inter-assay repeatability (4 plates) calculated for 1 positive and 4 negative controls was under 20%. The performance of this ELISA suggests that it could be used as a reliable and practical tool for seroepidemiological studies in Argentina.
publishDate 2021
dc.date.none.fl_str_mv 2021
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Development and validation of an Enzyme-Linked Immunosorbent Assay for the detection of antibodies against Trichinella spp. in domestic pigs in Argentina; 28th International Conference of the World Association for the Advancement of Veterinary Parasitology; Dublin; Reino Unido; 2021; 452-452
CONICET Digital
CONICET
url http://hdl.handle.net/11336/156172
identifier_str_mv Development and validation of an Enzyme-Linked Immunosorbent Assay for the detection of antibodies against Trichinella spp. in domestic pigs in Argentina; 28th International Conference of the World Association for the Advancement of Veterinary Parasitology; Dublin; Reino Unido; 2021; 452-452
CONICET Digital
CONICET
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dc.publisher.none.fl_str_mv World Association for the Advancement of Veterinary Parasitology
publisher.none.fl_str_mv World Association for the Advancement of Veterinary Parasitology
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