Development of a low-cost copro-LAMP assay for simultaneous copro-detection of Toxocara canis and Toxocara cati

Autores
Avila, Héctor Gabriel; Risso, Marikena Guadalupe; Ruybal, Paula; Repetto, Silvia; Butti, Marcos Javier; Trangoni, Marcos David; Grune Loffler, Sylvia; Perez, Veronica Mirtha; Periago, Maria Victoria
Año de publicación
2021
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
Toxocariasis is a zoonotic disease caused mainly by Toxocara canis and Toxocara cati and diagnosis in dogs and cats is an important tool for its control. For this reason, a new coprological loop-mediated isothermal amplification (LAMP) assay was developed for the simultaneous detection of these species. The primer set was designed on a region of the mitochondrial cox-1 gene. Amplification conditions were evaluated using a temperature gradient (52°C to 68°C), different incubation times (15-120 min), and different concentrations of malachite green dye (0.004-0.4% w/v). The analytical sensitivity was evaluated with serial dilutions of genomic DNA from T. canis and T. cati adult worms, and with serial dilutions of DNA extracted from feces using a low-cost in-house method. The specificity was evaluated using genomic DNA from Canis lupus familiaris, Felis catus, Escherichia coli, Toxascaris leonina, Ancylostoma caninum, Echinococcus granulosus sensu stricto and Taenia hydatigena. The LAMP assay applied to environmental fecal samples from an endemic area showed an analytical sensitivity of 10-100 fg of genomic DNA and 10-5 serial dilutions of DNA extracted from feces using the low-cost in-house method; with a specificity of 100%. Additionally, the total development of the assay was carried out in a basic laboratory and per-reaction reagent cost decreased by ~80%. This new, low-cost tool can help identify the most common agents of toxocariasis in endemic areas in order to manage prevention strategies without having to rely on a laboratory with sophisticated equipment.
Fil: Avila, Héctor Gabriel. Universidad Católica de Cuyo. Facultad de Ciencias Químicas y Tecnológicas. Laboratorio Provincial de Zoonosis Provincial; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - San Luis; Argentina
Fil: Risso, Marikena Guadalupe. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Investigaciones en Microbiología y Parasitología Médica. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Investigaciones en Microbiología y Parasitología Médica; Argentina
Fil: Ruybal, Paula. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Investigaciones en Microbiología y Parasitología Médica. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Investigaciones en Microbiología y Parasitología Médica; Argentina
Fil: Repetto, Silvia. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Investigaciones en Microbiología y Parasitología Médica. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Investigaciones en Microbiología y Parasitología Médica; Argentina
Fil: Butti, Marcos Javier. Universidad Nacional de La Plata. Facultad de Ciencias Veterinarias. Departamento de Microbiología. Laboratorio de Parasitosis Humanas y Zoonosis Parasitarias; Argentina
Fil: Trangoni, Marcos David. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Biotecnología; Argentina
Fil: Grune Loffler, Sylvia. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Patobiología Veterinaria - Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Patobiología Veterinaria; Argentina
Fil: Perez, Veronica Mirtha. Gobierno de la Provincia de San Juan. Ministerio de Salud Publica. Direccion de Epidemiologia. Seccion Rabia y Zoonosis.; Argentina
Fil: Periago, Maria Victoria. Fundación Mundo Sano; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Materia
COPRO-DIAGNOSIS
LOOP-MEDIATED ISOTHERMAL AMPLIFICATION
LOW-COST
TOXOCARA CANIS
TOXOCARA CATI
TOXOCARIASIS
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:11336/151870
Acceder
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oai_identifier_str oai:ri.conicet.gov.ar:11336/151870
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling Development of a low-cost copro-LAMP assay for simultaneous copro-detection of Toxocara canis and Toxocara catiAvila, Héctor GabrielRisso, Marikena GuadalupeRuybal, PaulaRepetto, SilviaButti, Marcos JavierTrangoni, Marcos DavidGrune Loffler, SylviaPerez, Veronica MirthaPeriago, Maria VictoriaCOPRO-DIAGNOSISLOOP-MEDIATED ISOTHERMAL AMPLIFICATIONLOW-COSTTOXOCARA CANISTOXOCARA CATITOXOCARIASIShttps://purl.org/becyt/ford/1.6https://purl.org/becyt/ford/1Toxocariasis is a zoonotic disease caused mainly by Toxocara canis and Toxocara cati and diagnosis in dogs and cats is an important tool for its control. For this reason, a new coprological loop-mediated isothermal amplification (LAMP) assay was developed for the simultaneous detection of these species. The primer set was designed on a region of the mitochondrial cox-1 gene. Amplification conditions were evaluated using a temperature gradient (52°C to 68°C), different incubation times (15-120 min), and different concentrations of malachite green dye (0.004-0.4% w/v). The analytical sensitivity was evaluated with serial dilutions of genomic DNA from T. canis and T. cati adult worms, and with serial dilutions of DNA extracted from feces using a low-cost in-house method. The specificity was evaluated using genomic DNA from Canis lupus familiaris, Felis catus, Escherichia coli, Toxascaris leonina, Ancylostoma caninum, Echinococcus granulosus sensu stricto and Taenia hydatigena. The LAMP assay applied to environmental fecal samples from an endemic area showed an analytical sensitivity of 10-100 fg of genomic DNA and 10-5 serial dilutions of DNA extracted from feces using the low-cost in-house method; with a specificity of 100%. Additionally, the total development of the assay was carried out in a basic laboratory and per-reaction reagent cost decreased by ~80%. This new, low-cost tool can help identify the most common agents of toxocariasis in endemic areas in order to manage prevention strategies without having to rely on a laboratory with sophisticated equipment.Fil: Avila, Héctor Gabriel. Universidad Católica de Cuyo. Facultad de Ciencias Químicas y Tecnológicas. Laboratorio Provincial de Zoonosis Provincial; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - San Luis; ArgentinaFil: Risso, Marikena Guadalupe. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Investigaciones en Microbiología y Parasitología Médica. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Investigaciones en Microbiología y Parasitología Médica; ArgentinaFil: Ruybal, Paula. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Investigaciones en Microbiología y Parasitología Médica. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Investigaciones en Microbiología y Parasitología Médica; ArgentinaFil: Repetto, Silvia. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Investigaciones en Microbiología y Parasitología Médica. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Investigaciones en Microbiología y Parasitología Médica; ArgentinaFil: Butti, Marcos Javier. Universidad Nacional de La Plata. Facultad de Ciencias Veterinarias. Departamento de Microbiología. Laboratorio de Parasitosis Humanas y Zoonosis Parasitarias; ArgentinaFil: Trangoni, Marcos David. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Biotecnología; ArgentinaFil: Grune Loffler, Sylvia. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Patobiología Veterinaria - Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Patobiología Veterinaria; ArgentinaFil: Perez, Veronica Mirtha. Gobierno de la Provincia de San Juan. Ministerio de Salud Publica. Direccion de Epidemiologia. Seccion Rabia y Zoonosis.; ArgentinaFil: Periago, Maria Victoria. Fundación Mundo Sano; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaCambridge University Press2021-06info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfapplication/pdfapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/151870Avila, Héctor Gabriel; Risso, Marikena Guadalupe; Ruybal, Paula; Repetto, Silvia; Butti, Marcos Javier; et al.; Development of a low-cost copro-LAMP assay for simultaneous copro-detection of Toxocara canis and Toxocara cati; Cambridge University Press; Parasitology; 148; 7; 6-2021; 819-8260031-18201469-8161CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/url/https://www.cambridge.org/core/product/identifier/S0031182021000342/type/journal_articleinfo:eu-repo/semantics/altIdentifier/doi/10.1017/S0031182021000342info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-09-03T09:51:07Zoai:ri.conicet.gov.ar:11336/151870instacron: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:34982025-09-03 09:51:07.705CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Development of a low-cost copro-LAMP assay for simultaneous copro-detection of Toxocara canis and Toxocara cati
title Development of a low-cost copro-LAMP assay for simultaneous copro-detection of Toxocara canis and Toxocara cati
spellingShingle Development of a low-cost copro-LAMP assay for simultaneous copro-detection of Toxocara canis and Toxocara cati
Avila, Héctor Gabriel
COPRO-DIAGNOSIS
LOOP-MEDIATED ISOTHERMAL AMPLIFICATION
LOW-COST
TOXOCARA CANIS
TOXOCARA CATI
TOXOCARIASIS
title_short Development of a low-cost copro-LAMP assay for simultaneous copro-detection of Toxocara canis and Toxocara cati
title_full Development of a low-cost copro-LAMP assay for simultaneous copro-detection of Toxocara canis and Toxocara cati
title_fullStr Development of a low-cost copro-LAMP assay for simultaneous copro-detection of Toxocara canis and Toxocara cati
title_full_unstemmed Development of a low-cost copro-LAMP assay for simultaneous copro-detection of Toxocara canis and Toxocara cati
title_sort Development of a low-cost copro-LAMP assay for simultaneous copro-detection of Toxocara canis and Toxocara cati
dc.creator.none.fl_str_mv Avila, Héctor Gabriel
Risso, Marikena Guadalupe
Ruybal, Paula
Repetto, Silvia
Butti, Marcos Javier
Trangoni, Marcos David
Grune Loffler, Sylvia
Perez, Veronica Mirtha
Periago, Maria Victoria
author Avila, Héctor Gabriel
author_facet Avila, Héctor Gabriel
Risso, Marikena Guadalupe
Ruybal, Paula
Repetto, Silvia
Butti, Marcos Javier
Trangoni, Marcos David
Grune Loffler, Sylvia
Perez, Veronica Mirtha
Periago, Maria Victoria
author_role author
author2 Risso, Marikena Guadalupe
Ruybal, Paula
Repetto, Silvia
Butti, Marcos Javier
Trangoni, Marcos David
Grune Loffler, Sylvia
Perez, Veronica Mirtha
Periago, Maria Victoria
author2_role author
author
author
author
author
author
author
author
dc.subject.none.fl_str_mv COPRO-DIAGNOSIS
LOOP-MEDIATED ISOTHERMAL AMPLIFICATION
LOW-COST
TOXOCARA CANIS
TOXOCARA CATI
TOXOCARIASIS
topic COPRO-DIAGNOSIS
LOOP-MEDIATED ISOTHERMAL AMPLIFICATION
LOW-COST
TOXOCARA CANIS
TOXOCARA CATI
TOXOCARIASIS
purl_subject.fl_str_mv https://purl.org/becyt/ford/1.6
https://purl.org/becyt/ford/1
dc.description.none.fl_txt_mv Toxocariasis is a zoonotic disease caused mainly by Toxocara canis and Toxocara cati and diagnosis in dogs and cats is an important tool for its control. For this reason, a new coprological loop-mediated isothermal amplification (LAMP) assay was developed for the simultaneous detection of these species. The primer set was designed on a region of the mitochondrial cox-1 gene. Amplification conditions were evaluated using a temperature gradient (52°C to 68°C), different incubation times (15-120 min), and different concentrations of malachite green dye (0.004-0.4% w/v). The analytical sensitivity was evaluated with serial dilutions of genomic DNA from T. canis and T. cati adult worms, and with serial dilutions of DNA extracted from feces using a low-cost in-house method. The specificity was evaluated using genomic DNA from Canis lupus familiaris, Felis catus, Escherichia coli, Toxascaris leonina, Ancylostoma caninum, Echinococcus granulosus sensu stricto and Taenia hydatigena. The LAMP assay applied to environmental fecal samples from an endemic area showed an analytical sensitivity of 10-100 fg of genomic DNA and 10-5 serial dilutions of DNA extracted from feces using the low-cost in-house method; with a specificity of 100%. Additionally, the total development of the assay was carried out in a basic laboratory and per-reaction reagent cost decreased by ~80%. This new, low-cost tool can help identify the most common agents of toxocariasis in endemic areas in order to manage prevention strategies without having to rely on a laboratory with sophisticated equipment.
Fil: Avila, Héctor Gabriel. Universidad Católica de Cuyo. Facultad de Ciencias Químicas y Tecnológicas. Laboratorio Provincial de Zoonosis Provincial; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - San Luis; Argentina
Fil: Risso, Marikena Guadalupe. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Investigaciones en Microbiología y Parasitología Médica. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Investigaciones en Microbiología y Parasitología Médica; Argentina
Fil: Ruybal, Paula. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Investigaciones en Microbiología y Parasitología Médica. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Investigaciones en Microbiología y Parasitología Médica; Argentina
Fil: Repetto, Silvia. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Investigaciones en Microbiología y Parasitología Médica. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Investigaciones en Microbiología y Parasitología Médica; Argentina
Fil: Butti, Marcos Javier. Universidad Nacional de La Plata. Facultad de Ciencias Veterinarias. Departamento de Microbiología. Laboratorio de Parasitosis Humanas y Zoonosis Parasitarias; Argentina
Fil: Trangoni, Marcos David. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Biotecnología; Argentina
Fil: Grune Loffler, Sylvia. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Patobiología Veterinaria - Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Patobiología Veterinaria; Argentina
Fil: Perez, Veronica Mirtha. Gobierno de la Provincia de San Juan. Ministerio de Salud Publica. Direccion de Epidemiologia. Seccion Rabia y Zoonosis.; Argentina
Fil: Periago, Maria Victoria. Fundación Mundo Sano; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
description Toxocariasis is a zoonotic disease caused mainly by Toxocara canis and Toxocara cati and diagnosis in dogs and cats is an important tool for its control. For this reason, a new coprological loop-mediated isothermal amplification (LAMP) assay was developed for the simultaneous detection of these species. The primer set was designed on a region of the mitochondrial cox-1 gene. Amplification conditions were evaluated using a temperature gradient (52°C to 68°C), different incubation times (15-120 min), and different concentrations of malachite green dye (0.004-0.4% w/v). The analytical sensitivity was evaluated with serial dilutions of genomic DNA from T. canis and T. cati adult worms, and with serial dilutions of DNA extracted from feces using a low-cost in-house method. The specificity was evaluated using genomic DNA from Canis lupus familiaris, Felis catus, Escherichia coli, Toxascaris leonina, Ancylostoma caninum, Echinococcus granulosus sensu stricto and Taenia hydatigena. The LAMP assay applied to environmental fecal samples from an endemic area showed an analytical sensitivity of 10-100 fg of genomic DNA and 10-5 serial dilutions of DNA extracted from feces using the low-cost in-house method; with a specificity of 100%. Additionally, the total development of the assay was carried out in a basic laboratory and per-reaction reagent cost decreased by ~80%. This new, low-cost tool can help identify the most common agents of toxocariasis in endemic areas in order to manage prevention strategies without having to rely on a laboratory with sophisticated equipment.
publishDate 2021
dc.date.none.fl_str_mv 2021-06
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/11336/151870
Avila, Héctor Gabriel; Risso, Marikena Guadalupe; Ruybal, Paula; Repetto, Silvia; Butti, Marcos Javier; et al.; Development of a low-cost copro-LAMP assay for simultaneous copro-detection of Toxocara canis and Toxocara cati; Cambridge University Press; Parasitology; 148; 7; 6-2021; 819-826
0031-1820
1469-8161
CONICET Digital
CONICET
url http://hdl.handle.net/11336/151870
identifier_str_mv Avila, Héctor Gabriel; Risso, Marikena Guadalupe; Ruybal, Paula; Repetto, Silvia; Butti, Marcos Javier; et al.; Development of a low-cost copro-LAMP assay for simultaneous copro-detection of Toxocara canis and Toxocara cati; Cambridge University Press; Parasitology; 148; 7; 6-2021; 819-826
0031-1820
1469-8161
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://www.cambridge.org/core/product/identifier/S0031182021000342/type/journal_article
info:eu-repo/semantics/altIdentifier/doi/10.1017/S0031182021000342
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
application/pdf
application/pdf
dc.publisher.none.fl_str_mv Cambridge University Press
publisher.none.fl_str_mv Cambridge University Press
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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