Development and standardization of a Loop-mediated isothermal amplification (LAMP) test for the detection of Babesia bigemina

Autores
Lizarazo Zuluaga, Andrea P.; Carvajal Gamez, Bertha I.; Wilkowsky, Silvina Elizabeth; Cravero, Silvio Lorenzo Pedro; Trangoni, Marcos David; Mosqueda, Juan
Año de publicación
2022
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
Bovine babesiosis is a tick-borne disease caused by protozoan parasites of the genus Babesia. Babesia bigemina is one of the most prevalent and economically important parasite species that infects cattle because of its impact on the meat and milk production industry. Effective disease control strategies should include detection of reservoir animals and early and specific pathogen detection using rapid, economical, sensitive, and specific detection techniques. The loop-mediated isothermal amplification technique (LAMP) is a one-step molecular reaction that amplifies DNA sequences with high sensitivity and specificity under isothermal conditions and requires no special equipment. The results can be observed by the naked eye as color changes. The aim of this work was to develop and standardize the LAMP technique for B. bigemina detection and its visualization using hydroxynaphtol blue. For this situation, primers were designed from the conserved sequences of the B. bigemina ama-1 gene. The results showed that at 63 °C in 1 h and under standardized conditions, this technique could amplify B. bigemina DNA as indicated by the characteristic colorimetric change. Sensitivity evaluation indicated that DNA was amplified at a 0.00000001% parasitemia, and it was demonstrated that this technique specifically amplified the DNA of B. bigemina. Additionally, this technique could amplify DNA from 10 strains of B. bigemina from three different countries. It is concluded that the LAMP technique as modified in our case could specifically amplify B. bigemina DNA and shows high sensitivity, does not cross-react with related organisms, and the product is observed by 60 min of reaction time based on color changes. This report is the first LAMP report that uses sequences that are conserved between strains of the ama-1 gene, demonstrates the results by color changes using hydroxynaphtol blue. We propose LAMP as a rapid and economical alternative method for the molecular detection of B. bigemina.
Fil: Lizarazo Zuluaga, Andrea P.. Universidad Autonoma de Queretaro.; México
Fil: Carvajal Gamez, Bertha I.. Universidad Autonoma de Queretaro.; México
Fil: Wilkowsky, Silvina Elizabeth. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Agrobiotecnología y Biología Molecular. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Agrobiotecnología y Biología Molecular; Argentina
Fil: Cravero, Silvio Lorenzo Pedro. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Agrobiotecnología y Biología Molecular. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Agrobiotecnología y Biología Molecular; Argentina
Fil: Trangoni, Marcos David. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Agrobiotecnología y Biología Molecular. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Agrobiotecnología y Biología Molecular; Argentina
Fil: Mosqueda, Juan. Universidad Autonoma de Queretaro.; México
Materia
BABESIA BIGEMINA
DIAGNOSTICS
HYDROXYNAPHTOL BLUE
LAMP
MOLECULAR DETECTION
Nivel de accesibilidad
acceso abierto
Condiciones de uso
https://creativecommons.org/licenses/by/2.5/ar/
Repositorio
CONICET Digital (CONICET)
Institución
Consejo Nacional de Investigaciones Científicas y Técnicas
OAI Identificador
oai:ri.conicet.gov.ar:11336/213473

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spelling Development and standardization of a Loop-mediated isothermal amplification (LAMP) test for the detection of Babesia bigeminaLizarazo Zuluaga, Andrea P.Carvajal Gamez, Bertha I.Wilkowsky, Silvina ElizabethCravero, Silvio Lorenzo PedroTrangoni, Marcos DavidMosqueda, JuanBABESIA BIGEMINADIAGNOSTICSHYDROXYNAPHTOL BLUELAMPMOLECULAR DETECTIONhttps://purl.org/becyt/ford/1.6https://purl.org/becyt/ford/1Bovine babesiosis is a tick-borne disease caused by protozoan parasites of the genus Babesia. Babesia bigemina is one of the most prevalent and economically important parasite species that infects cattle because of its impact on the meat and milk production industry. Effective disease control strategies should include detection of reservoir animals and early and specific pathogen detection using rapid, economical, sensitive, and specific detection techniques. The loop-mediated isothermal amplification technique (LAMP) is a one-step molecular reaction that amplifies DNA sequences with high sensitivity and specificity under isothermal conditions and requires no special equipment. The results can be observed by the naked eye as color changes. The aim of this work was to develop and standardize the LAMP technique for B. bigemina detection and its visualization using hydroxynaphtol blue. For this situation, primers were designed from the conserved sequences of the B. bigemina ama-1 gene. The results showed that at 63 °C in 1 h and under standardized conditions, this technique could amplify B. bigemina DNA as indicated by the characteristic colorimetric change. Sensitivity evaluation indicated that DNA was amplified at a 0.00000001% parasitemia, and it was demonstrated that this technique specifically amplified the DNA of B. bigemina. Additionally, this technique could amplify DNA from 10 strains of B. bigemina from three different countries. It is concluded that the LAMP technique as modified in our case could specifically amplify B. bigemina DNA and shows high sensitivity, does not cross-react with related organisms, and the product is observed by 60 min of reaction time based on color changes. This report is the first LAMP report that uses sequences that are conserved between strains of the ama-1 gene, demonstrates the results by color changes using hydroxynaphtol blue. We propose LAMP as a rapid and economical alternative method for the molecular detection of B. bigemina.Fil: Lizarazo Zuluaga, Andrea P.. Universidad Autonoma de Queretaro.; MéxicoFil: Carvajal Gamez, Bertha I.. Universidad Autonoma de Queretaro.; MéxicoFil: Wilkowsky, Silvina Elizabeth. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Agrobiotecnología y Biología Molecular. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Agrobiotecnología y Biología Molecular; ArgentinaFil: Cravero, Silvio Lorenzo Pedro. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Agrobiotecnología y Biología Molecular. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Agrobiotecnología y Biología Molecular; ArgentinaFil: Trangoni, Marcos David. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Agrobiotecnología y Biología Molecular. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Agrobiotecnología y Biología Molecular; ArgentinaFil: Mosqueda, Juan. Universidad Autonoma de Queretaro.; MéxicoFrontiers Media2022-11info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/213473Lizarazo Zuluaga, Andrea P.; Carvajal Gamez, Bertha I.; Wilkowsky, Silvina Elizabeth; Cravero, Silvio Lorenzo Pedro; Trangoni, Marcos David; et al.; Development and standardization of a Loop-mediated isothermal amplification (LAMP) test for the detection of Babesia bigemina; Frontiers Media; Frontiers in Veterinary Science; 9; 11-2022; 1-102297-1769CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/url/https://www.frontiersin.org/articles/10.3389/fvets.2022.1056355/fullinfo:eu-repo/semantics/altIdentifier/doi/10.3389/fvets.2022.1056355info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-09-29T10:42:30Zoai:ri.conicet.gov.ar:11336/213473instacron: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 10:42:31.211CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Development and standardization of a Loop-mediated isothermal amplification (LAMP) test for the detection of Babesia bigemina
title Development and standardization of a Loop-mediated isothermal amplification (LAMP) test for the detection of Babesia bigemina
spellingShingle Development and standardization of a Loop-mediated isothermal amplification (LAMP) test for the detection of Babesia bigemina
Lizarazo Zuluaga, Andrea P.
BABESIA BIGEMINA
DIAGNOSTICS
HYDROXYNAPHTOL BLUE
LAMP
MOLECULAR DETECTION
title_short Development and standardization of a Loop-mediated isothermal amplification (LAMP) test for the detection of Babesia bigemina
title_full Development and standardization of a Loop-mediated isothermal amplification (LAMP) test for the detection of Babesia bigemina
title_fullStr Development and standardization of a Loop-mediated isothermal amplification (LAMP) test for the detection of Babesia bigemina
title_full_unstemmed Development and standardization of a Loop-mediated isothermal amplification (LAMP) test for the detection of Babesia bigemina
title_sort Development and standardization of a Loop-mediated isothermal amplification (LAMP) test for the detection of Babesia bigemina
dc.creator.none.fl_str_mv Lizarazo Zuluaga, Andrea P.
Carvajal Gamez, Bertha I.
Wilkowsky, Silvina Elizabeth
Cravero, Silvio Lorenzo Pedro
Trangoni, Marcos David
Mosqueda, Juan
author Lizarazo Zuluaga, Andrea P.
author_facet Lizarazo Zuluaga, Andrea P.
Carvajal Gamez, Bertha I.
Wilkowsky, Silvina Elizabeth
Cravero, Silvio Lorenzo Pedro
Trangoni, Marcos David
Mosqueda, Juan
author_role author
author2 Carvajal Gamez, Bertha I.
Wilkowsky, Silvina Elizabeth
Cravero, Silvio Lorenzo Pedro
Trangoni, Marcos David
Mosqueda, Juan
author2_role author
author
author
author
author
dc.subject.none.fl_str_mv BABESIA BIGEMINA
DIAGNOSTICS
HYDROXYNAPHTOL BLUE
LAMP
MOLECULAR DETECTION
topic BABESIA BIGEMINA
DIAGNOSTICS
HYDROXYNAPHTOL BLUE
LAMP
MOLECULAR DETECTION
purl_subject.fl_str_mv https://purl.org/becyt/ford/1.6
https://purl.org/becyt/ford/1
dc.description.none.fl_txt_mv Bovine babesiosis is a tick-borne disease caused by protozoan parasites of the genus Babesia. Babesia bigemina is one of the most prevalent and economically important parasite species that infects cattle because of its impact on the meat and milk production industry. Effective disease control strategies should include detection of reservoir animals and early and specific pathogen detection using rapid, economical, sensitive, and specific detection techniques. The loop-mediated isothermal amplification technique (LAMP) is a one-step molecular reaction that amplifies DNA sequences with high sensitivity and specificity under isothermal conditions and requires no special equipment. The results can be observed by the naked eye as color changes. The aim of this work was to develop and standardize the LAMP technique for B. bigemina detection and its visualization using hydroxynaphtol blue. For this situation, primers were designed from the conserved sequences of the B. bigemina ama-1 gene. The results showed that at 63 °C in 1 h and under standardized conditions, this technique could amplify B. bigemina DNA as indicated by the characteristic colorimetric change. Sensitivity evaluation indicated that DNA was amplified at a 0.00000001% parasitemia, and it was demonstrated that this technique specifically amplified the DNA of B. bigemina. Additionally, this technique could amplify DNA from 10 strains of B. bigemina from three different countries. It is concluded that the LAMP technique as modified in our case could specifically amplify B. bigemina DNA and shows high sensitivity, does not cross-react with related organisms, and the product is observed by 60 min of reaction time based on color changes. This report is the first LAMP report that uses sequences that are conserved between strains of the ama-1 gene, demonstrates the results by color changes using hydroxynaphtol blue. We propose LAMP as a rapid and economical alternative method for the molecular detection of B. bigemina.
Fil: Lizarazo Zuluaga, Andrea P.. Universidad Autonoma de Queretaro.; México
Fil: Carvajal Gamez, Bertha I.. Universidad Autonoma de Queretaro.; México
Fil: Wilkowsky, Silvina Elizabeth. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Agrobiotecnología y Biología Molecular. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Agrobiotecnología y Biología Molecular; Argentina
Fil: Cravero, Silvio Lorenzo Pedro. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Agrobiotecnología y Biología Molecular. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Agrobiotecnología y Biología Molecular; Argentina
Fil: Trangoni, Marcos David. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Agrobiotecnología y Biología Molecular. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Agrobiotecnología y Biología Molecular; Argentina
Fil: Mosqueda, Juan. Universidad Autonoma de Queretaro.; México
description Bovine babesiosis is a tick-borne disease caused by protozoan parasites of the genus Babesia. Babesia bigemina is one of the most prevalent and economically important parasite species that infects cattle because of its impact on the meat and milk production industry. Effective disease control strategies should include detection of reservoir animals and early and specific pathogen detection using rapid, economical, sensitive, and specific detection techniques. The loop-mediated isothermal amplification technique (LAMP) is a one-step molecular reaction that amplifies DNA sequences with high sensitivity and specificity under isothermal conditions and requires no special equipment. The results can be observed by the naked eye as color changes. The aim of this work was to develop and standardize the LAMP technique for B. bigemina detection and its visualization using hydroxynaphtol blue. For this situation, primers were designed from the conserved sequences of the B. bigemina ama-1 gene. The results showed that at 63 °C in 1 h and under standardized conditions, this technique could amplify B. bigemina DNA as indicated by the characteristic colorimetric change. Sensitivity evaluation indicated that DNA was amplified at a 0.00000001% parasitemia, and it was demonstrated that this technique specifically amplified the DNA of B. bigemina. Additionally, this technique could amplify DNA from 10 strains of B. bigemina from three different countries. It is concluded that the LAMP technique as modified in our case could specifically amplify B. bigemina DNA and shows high sensitivity, does not cross-react with related organisms, and the product is observed by 60 min of reaction time based on color changes. This report is the first LAMP report that uses sequences that are conserved between strains of the ama-1 gene, demonstrates the results by color changes using hydroxynaphtol blue. We propose LAMP as a rapid and economical alternative method for the molecular detection of B. bigemina.
publishDate 2022
dc.date.none.fl_str_mv 2022-11
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/11336/213473
Lizarazo Zuluaga, Andrea P.; Carvajal Gamez, Bertha I.; Wilkowsky, Silvina Elizabeth; Cravero, Silvio Lorenzo Pedro; Trangoni, Marcos David; et al.; Development and standardization of a Loop-mediated isothermal amplification (LAMP) test for the detection of Babesia bigemina; Frontiers Media; Frontiers in Veterinary Science; 9; 11-2022; 1-10
2297-1769
CONICET Digital
CONICET
url http://hdl.handle.net/11336/213473
identifier_str_mv Lizarazo Zuluaga, Andrea P.; Carvajal Gamez, Bertha I.; Wilkowsky, Silvina Elizabeth; Cravero, Silvio Lorenzo Pedro; Trangoni, Marcos David; et al.; Development and standardization of a Loop-mediated isothermal amplification (LAMP) test for the detection of Babesia bigemina; Frontiers Media; Frontiers in Veterinary Science; 9; 11-2022; 1-10
2297-1769
CONICET Digital
CONICET
dc.language.none.fl_str_mv eng
language eng
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info:eu-repo/semantics/altIdentifier/doi/10.3389/fvets.2022.1056355
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
https://creativecommons.org/licenses/by/2.5/ar/
eu_rights_str_mv openAccess
rights_invalid_str_mv https://creativecommons.org/licenses/by/2.5/ar/
dc.format.none.fl_str_mv application/pdf
application/pdf
dc.publisher.none.fl_str_mv Frontiers Media
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dc.source.none.fl_str_mv reponame:CONICET Digital (CONICET)
instname:Consejo Nacional de Investigaciones Científicas y Técnicas
reponame_str CONICET Digital (CONICET)
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repository.mail.fl_str_mv dasensio@conicet.gov.ar; lcarlino@conicet.gov.ar
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