Loop-mediated isothermal amplification (LAMP) assay for the diagnosis of fasciolosis in sheep and its application under field conditions

[Background] Loop-mediated isothermal amplification (LAMP) is a very specific, efficient, and rapid gene amplification procedure in which the reaction can run at a constant temperature. In the current study we have developed a LAMP assay to improve the diagnosis of Fasciola spp. in the faeces of she...

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Detalhes bibliográficos
Autores: Martínez Valladares, María, Rojo Vázquez, Francisco Antonio
Formato: artículo
Estado:Versión publicada
Fecha de publicación:2016
País:España
Recursos:Consejo Superior de Investigaciones Científicas (CSIC)
Repositorio:DIGITAL.CSIC. Repositorio Institucional del CSIC
OAI Identifier:oai:digital.csic.es:10261/143361
Acesso em linha:http://hdl.handle.net/10261/143361
Access Level:acceso abierto
Palavra-chave:Fasciola spp
Sheep
Diagnosis
PCR
Loop-mediated isothermal amplification (LAMP)
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spelling Loop-mediated isothermal amplification (LAMP) assay for the diagnosis of fasciolosis in sheep and its application under field conditionsMartínez Valladares, MaríaRojo Vázquez, Francisco AntonioFasciola sppSheepDiagnosisPCRLoop-mediated isothermal amplification (LAMP)[Background] Loop-mediated isothermal amplification (LAMP) is a very specific, efficient, and rapid gene amplification procedure in which the reaction can run at a constant temperature. In the current study we have developed a LAMP assay to improve the diagnosis of Fasciola spp. in the faeces of sheep.[Findings] After the optimisation of the LAMP assay we have shown similar results between this technique and the standard PCR using the outer primers of the LAMP reaction. In both cases the limit of detection was 10 pg; also, the diagnosis of fasciolosis was confirmed during the first week post-infection in experimental infected sheep by both techniques. In eight naturally infected sheep, the infection with F. hepatica was confirmed in all animals before a treatment with triclabendazole and on day 30 post treatment in two sheep using the LAMP assay; however, when we carried out the standard PCR with the outer primers, the results before treatment were the same but on day 30 post-treatment the infection was only confirmed in one out of the two sheep. On the other hand, the standard PCR took around 3 h to obtain a result, comparing with 1 h and 10 min for the LAMP assay.[Conclusions] The LAMP assay described here could be a good alternative to conventional diagnostic methods to detect F. hepatica in faeces since it solves the drawbacks of the standard PCR.We thank Drs Manga-González and González-Lanza (Instituto de Ganadería de Montaña, CSIC-ULE, Spain) for providing DNA samples of D. dendriticum and C. daubneyi. This study has been funded by the national project INIA (Instituto Nacional de Investigaciones Agrarias: RTA2010-00094-C03-02), Ministry of Economy and Competitiveness (Ministerio de Economia y Competitividad), and also co-funded by the European Regional Development Fund (Fondos Feder).We acknowledge support by the CSIC Open Access Publication Initiative through its Unit of Information Resources for Research (URICI).Peer reviewedBioMed CentralCSIC - Instituto Nacional de Investigación y Tecnología Agraria y Alimentaria (INIA)Ministerio de Economía y Competitividad (España)European CommissionCSIC - Unidad de Recursos de Información Científica para la Investigación (URICI)Consejo Superior de Investigaciones Científicas [https://ror.org/02gfc7t72]201720172016info:eu-repo/semantics/articlehttp://purl.org/coar/resource_type/c_6501Publisher's versioninfo:eu-repo/semantics/publishedVersionhttp://hdl.handle.net/10261/143361reponame:DIGITAL.CSIC. Repositorio Institucional del CSICinstname:Consejo Superior de Investigaciones Científicas (CSIC)Ingléshttps://doi.org/10.1186/s13071-016-1355-2Síinfo:eu-repo/semantics/openAccessoai:digital.csic.es:10261/1433612026-05-22T06:33:51Z
dc.title.none.fl_str_mv Loop-mediated isothermal amplification (LAMP) assay for the diagnosis of fasciolosis in sheep and its application under field conditions
title Loop-mediated isothermal amplification (LAMP) assay for the diagnosis of fasciolosis in sheep and its application under field conditions
spellingShingle Loop-mediated isothermal amplification (LAMP) assay for the diagnosis of fasciolosis in sheep and its application under field conditions
Martínez Valladares, María
Fasciola spp
Sheep
Diagnosis
PCR
Loop-mediated isothermal amplification (LAMP)
title_short Loop-mediated isothermal amplification (LAMP) assay for the diagnosis of fasciolosis in sheep and its application under field conditions
title_full Loop-mediated isothermal amplification (LAMP) assay for the diagnosis of fasciolosis in sheep and its application under field conditions
title_fullStr Loop-mediated isothermal amplification (LAMP) assay for the diagnosis of fasciolosis in sheep and its application under field conditions
title_full_unstemmed Loop-mediated isothermal amplification (LAMP) assay for the diagnosis of fasciolosis in sheep and its application under field conditions
title_sort Loop-mediated isothermal amplification (LAMP) assay for the diagnosis of fasciolosis in sheep and its application under field conditions
dc.creator.none.fl_str_mv Martínez Valladares, María
Rojo Vázquez, Francisco Antonio
author Martínez Valladares, María
author_facet Martínez Valladares, María
Rojo Vázquez, Francisco Antonio
author_role author
author2 Rojo Vázquez, Francisco Antonio
author2_role author
dc.contributor.none.fl_str_mv CSIC - Instituto Nacional de Investigación y Tecnología Agraria y Alimentaria (INIA)
Ministerio de Economía y Competitividad (España)
European Commission
CSIC - Unidad de Recursos de Información Científica para la Investigación (URICI)
Consejo Superior de Investigaciones Científicas [https://ror.org/02gfc7t72]
dc.subject.none.fl_str_mv Fasciola spp
Sheep
Diagnosis
PCR
Loop-mediated isothermal amplification (LAMP)
topic Fasciola spp
Sheep
Diagnosis
PCR
Loop-mediated isothermal amplification (LAMP)
description [Background] Loop-mediated isothermal amplification (LAMP) is a very specific, efficient, and rapid gene amplification procedure in which the reaction can run at a constant temperature. In the current study we have developed a LAMP assay to improve the diagnosis of Fasciola spp. in the faeces of sheep.
publishDate 2016
dc.date.none.fl_str_mv 2016
2017
2017
dc.type.none.fl_str_mv info:eu-repo/semantics/article
http://purl.org/coar/resource_type/c_6501
Publisher's version
info:eu-repo/semantics/publishedVersion
format article
status_str publishedVersion
dc.identifier.none.fl_str_mv http://hdl.handle.net/10261/143361
url http://hdl.handle.net/10261/143361
dc.language.none.fl_str_mv Inglés
language_invalid_str_mv Inglés
dc.relation.none.fl_str_mv https://doi.org/10.1186/s13071-016-1355-2
Sí
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.publisher.none.fl_str_mv BioMed Central
publisher.none.fl_str_mv BioMed Central
dc.source.none.fl_str_mv reponame:DIGITAL.CSIC. Repositorio Institucional del CSIC
instname:Consejo Superior de Investigaciones Científicas (CSIC)
instname_str Consejo Superior de Investigaciones Científicas (CSIC)
reponame_str DIGITAL.CSIC. Repositorio Institucional del CSIC
collection DIGITAL.CSIC. Repositorio Institucional del CSIC
repository.name.fl_str_mv
repository.mail.fl_str_mv
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score 15,812429

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