Development of a real-time PCR assay for detection and quantification of Enteromyxum scophthalmi parasites in turbot intestinal samples

The myxozoan parasite Enteromyxum scophthalmi causes severe enteritis in cultured turbot Scophthalmus maximus, thus generating important economic losses. At present, there are no prevention or control measures for the disease, and many aspects of the life cycle and transmission of the parasite are n...

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Detalhes bibliográficos
Autores: Piazzon, Carla, Mallo Seijas, Natalia, Martín, Iris, Fernández Casal, Jacobo, Sanmartín, Manuel L., Lamas Fernández, Jesús, Leiro Vidal, José Manuel
Tipo de documento: artigo
Data de publicação:2012
País:España
Recursos:Universidad de Santiago de Compostela (USC)
Repositório:Minerva. Repositorio Institucional de la Universidad de Santiago de Compostela
Idioma:inglês
OAI Identifier:oai:minerva.usc.gal:10347/45920
Acesso em linha:https://hdl.handle.net/10347/45920
Access Level:Acceso aberto
Palavra-chave:Turbot
Enteromyxum scophthalmi
Myxozoan parasite
Quantitative PCR
Diagnosis
2414 Microbiología
240112 Parasitología animal
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spelling Development of a real-time PCR assay for detection and quantification of Enteromyxum scophthalmi parasites in turbot intestinal samplesPiazzon, CarlaMallo Seijas, NataliaMartín, IrisFernández Casal, JacoboSanmartín, Manuel L.Lamas Fernández, JesúsLeiro Vidal, José ManuelTurbotEnteromyxum scophthalmiMyxozoan parasiteQuantitative PCRDiagnosis2414 Microbiología240112 Parasitología animalThe myxozoan parasite Enteromyxum scophthalmi causes severe enteritis in cultured turbot Scophthalmus maximus, thus generating important economic losses. At present, there are no prevention or control measures for the disease, and many aspects of the life cycle and transmission of the parasite are not yet known. In this study, a highly sensitive, reproducible and rapid quantitative (real time) polymerase chain reaction (qPCR) assay was developed to detect E. scophthalmi DNA. The qPCR assay targets the 28S rRNA gene of the parasite, which has a high identity (94%) with the myxosporidian Enteromyxum leei rRNA gene. The qPCR assay was able to detect up to 13 DNA copies, corresponding to 0.55 fg, estimating that genomic DNA has around 1450 copies of 28S rRNA gene per parasite nucleus. The mean intra- and inter-assay coefficients of variation were below 5% and no detectable amplification was observed with DNA from non-infected turbot. The assay was validated with a histological identification of intestinal content samples from experimentally infected turbot and a good correlation between both methods was observed. The results demonstrate that the qPCR assay can be applied in the diagnosis of turbot enteromyxosis and to determine the relative abundance of E. scophthalmi in turbot intestinal contents in health monitoring studies.ElsevierUniversidade de Santiago de Compostela. Departamento de Bioloxía Celular e EcoloxíaUniversidade de Santiago de Compostela. Departamento de Microbioloxía e Parasitoloxía20122012-09-1520122012-09-15journal articlehttp://purl.org/coar/resource_type/c_6501AMhttp://purl.org/coar/version/c_ab4af688f83e57aainfo:eu-repo/semantics/articleapplication/pdfhttps://hdl.handle.net/10347/45920reponame:Minerva. Repositorio Institucional de la Universidad de Santiago de Compostelainstname:Universidad de Santiago de Compostela (USC)Inglésengopen accesshttp://purl.org/coar/access_right/c_abf2Attribution-NonCommercial-NoDerivatives 4.0 Internationalhttp://creativecommons.org/licenses/by-nc-nd/4.0/info:eu-repo/semantics/openAccessoai:minerva.usc.gal:10347/459202026-06-15T12:47:27Z
dc.title.none.fl_str_mv Development of a real-time PCR assay for detection and quantification of Enteromyxum scophthalmi parasites in turbot intestinal samples
title Development of a real-time PCR assay for detection and quantification of Enteromyxum scophthalmi parasites in turbot intestinal samples
spellingShingle Development of a real-time PCR assay for detection and quantification of Enteromyxum scophthalmi parasites in turbot intestinal samples
Piazzon, Carla
Turbot
Enteromyxum scophthalmi
Myxozoan parasite
Quantitative PCR
Diagnosis
2414 Microbiología
240112 Parasitología animal
title_short Development of a real-time PCR assay for detection and quantification of Enteromyxum scophthalmi parasites in turbot intestinal samples
title_full Development of a real-time PCR assay for detection and quantification of Enteromyxum scophthalmi parasites in turbot intestinal samples
title_fullStr Development of a real-time PCR assay for detection and quantification of Enteromyxum scophthalmi parasites in turbot intestinal samples
title_full_unstemmed Development of a real-time PCR assay for detection and quantification of Enteromyxum scophthalmi parasites in turbot intestinal samples
title_sort Development of a real-time PCR assay for detection and quantification of Enteromyxum scophthalmi parasites in turbot intestinal samples
dc.creator.none.fl_str_mv Piazzon, Carla
Mallo Seijas, Natalia
Martín, Iris
Fernández Casal, Jacobo
Sanmartín, Manuel L.
Lamas Fernández, Jesús
Leiro Vidal, José Manuel
author Piazzon, Carla
author_facet Piazzon, Carla
Mallo Seijas, Natalia
Martín, Iris
Fernández Casal, Jacobo
Sanmartín, Manuel L.
Lamas Fernández, Jesús
Leiro Vidal, José Manuel
author_role author
author2 Mallo Seijas, Natalia
Martín, Iris
Fernández Casal, Jacobo
Sanmartín, Manuel L.
Lamas Fernández, Jesús
Leiro Vidal, José Manuel
author2_role author
author
author
author
author
author
dc.contributor.none.fl_str_mv Universidade de Santiago de Compostela. Departamento de Bioloxía Celular e Ecoloxía
Universidade de Santiago de Compostela. Departamento de Microbioloxía e Parasitoloxía
dc.subject.none.fl_str_mv Turbot
Enteromyxum scophthalmi
Myxozoan parasite
Quantitative PCR
Diagnosis
2414 Microbiología
240112 Parasitología animal
topic Turbot
Enteromyxum scophthalmi
Myxozoan parasite
Quantitative PCR
Diagnosis
2414 Microbiología
240112 Parasitología animal
description The myxozoan parasite Enteromyxum scophthalmi causes severe enteritis in cultured turbot Scophthalmus maximus, thus generating important economic losses. At present, there are no prevention or control measures for the disease, and many aspects of the life cycle and transmission of the parasite are not yet known. In this study, a highly sensitive, reproducible and rapid quantitative (real time) polymerase chain reaction (qPCR) assay was developed to detect E. scophthalmi DNA. The qPCR assay targets the 28S rRNA gene of the parasite, which has a high identity (94%) with the myxosporidian Enteromyxum leei rRNA gene. The qPCR assay was able to detect up to 13 DNA copies, corresponding to 0.55 fg, estimating that genomic DNA has around 1450 copies of 28S rRNA gene per parasite nucleus. The mean intra- and inter-assay coefficients of variation were below 5% and no detectable amplification was observed with DNA from non-infected turbot. The assay was validated with a histological identification of intestinal content samples from experimentally infected turbot and a good correlation between both methods was observed. The results demonstrate that the qPCR assay can be applied in the diagnosis of turbot enteromyxosis and to determine the relative abundance of E. scophthalmi in turbot intestinal contents in health monitoring studies.
publishDate 2012
dc.date.none.fl_str_mv 2012
2012-09-15
2012
2012-09-15
dc.type.none.fl_str_mv journal article
http://purl.org/coar/resource_type/c_6501
AM
http://purl.org/coar/version/c_ab4af688f83e57aa
dc.type.openaire.fl_str_mv info:eu-repo/semantics/article
format article
dc.identifier.none.fl_str_mv https://hdl.handle.net/10347/45920
url https://hdl.handle.net/10347/45920
dc.language.none.fl_str_mv Inglés
eng
language_invalid_str_mv Inglés
language eng
dc.rights.none.fl_str_mv open access
http://purl.org/coar/access_right/c_abf2
Attribution-NonCommercial-NoDerivatives 4.0 International
http://creativecommons.org/licenses/by-nc-nd/4.0/
dc.rights.openaire.fl_str_mv info:eu-repo/semantics/openAccess
rights_invalid_str_mv open access
http://purl.org/coar/access_right/c_abf2
Attribution-NonCommercial-NoDerivatives 4.0 International
http://creativecommons.org/licenses/by-nc-nd/4.0/
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv application/pdf
dc.publisher.none.fl_str_mv Elsevier
publisher.none.fl_str_mv Elsevier
dc.source.none.fl_str_mv reponame:Minerva. Repositorio Institucional de la Universidad de Santiago de Compostela
instname:Universidad de Santiago de Compostela (USC)
instname_str Universidad de Santiago de Compostela (USC)
reponame_str Minerva. Repositorio Institucional de la Universidad de Santiago de Compostela
collection Minerva. Repositorio Institucional de la Universidad de Santiago de Compostela
repository.name.fl_str_mv
repository.mail.fl_str_mv
_version_ 1869411683672260608
score 15,228081

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