Satellite DNA as a target for TaqMan real-time PCR detection of the pinewood nematode, Bursaphelenchus xylophilus

Detalhes bibliográficos
Autor(a) principal: CECILE, FRANÇOIS
Data de Publicação: 2007
Outros Autores: CASTAGNONE, CHANTAL, BOONHAM, NEIL, TOMLINSON, JENNY, LAWSON, REBECCA, HOCKLAND, SUE, QUILL, JAMES, VIEIRA, Paulo, Mota, Manuel, CASTAGNONE-SERENO, PHILIPPE
Tipo de documento: Artigo
Idioma: eng
Título da fonte: Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
Texto Completo: http://hdl.handle.net/10174/6790
Resumo: The pinewood nematode (PWN), Bursaphelenchus xylophilus , is a major pathogen of conifers, which impacts on forest health, natural ecosystem stability and international trade. As a consequence, it has been listed as a quarantine organism in Europe. A real-time PCR approach based on TaqMan chemistry was developed to detect this organism. Specific probe and primers were designed based on the sequence of the Msp I satellite DNA family previously characterized in the genome of the nematode. The method proved to be specific in tests with target DNA from PWN isolates from worldwide origin. From a practical point of view, detection limit was 1 pg of target DNA or one individual nematode. In addition, PWN genomic DNA or single individuals were positively detected in mixed samples in which B. xylophilius was associated with the closely related non-pathogenic species B. mucronatus , up to the limit of 0.01% or 1% of the mixture, respectively. The real-time PCR assay was also used in conjunction with a simple DNA extraction method to detect PWN directly in artificially infested wood samples. These results demonstrate the potential of this assay to provide rapid, accurate and sensitive molecular identification of the PWN in relation to pest risk assessment in the field and quarantine regulation.
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spelling Satellite DNA as a target for TaqMan real-time PCR detection of the pinewood nematode, Bursaphelenchus xylophilusBursaphelenchus xylophiluspinewoodnematodeThe pinewood nematode (PWN), Bursaphelenchus xylophilus , is a major pathogen of conifers, which impacts on forest health, natural ecosystem stability and international trade. As a consequence, it has been listed as a quarantine organism in Europe. A real-time PCR approach based on TaqMan chemistry was developed to detect this organism. Specific probe and primers were designed based on the sequence of the Msp I satellite DNA family previously characterized in the genome of the nematode. The method proved to be specific in tests with target DNA from PWN isolates from worldwide origin. From a practical point of view, detection limit was 1 pg of target DNA or one individual nematode. In addition, PWN genomic DNA or single individuals were positively detected in mixed samples in which B. xylophilius was associated with the closely related non-pathogenic species B. mucronatus , up to the limit of 0.01% or 1% of the mixture, respectively. The real-time PCR assay was also used in conjunction with a simple DNA extraction method to detect PWN directly in artificially infested wood samples. These results demonstrate the potential of this assay to provide rapid, accurate and sensitive molecular identification of the PWN in relation to pest risk assessment in the field and quarantine regulation.2012-12-10T16:25:43Z2012-12-102007-01-01T00:00:00Zinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articlehttp://hdl.handle.net/10174/6790http://hdl.handle.net/10174/6790eng8ICAAMndndndndndndndpvieira@uevora.ptmmtota@uevora.ptPhilippe.Castagnone@sophia.inra.fr218CECILE, FRANÇOISCASTAGNONE, CHANTALBOONHAM, NEILTOMLINSON, JENNYLAWSON, REBECCAHOCKLAND, SUEQUILL, JAMESVIEIRA, PauloMota, ManuelCASTAGNONE-SERENO, PHILIPPEinfo:eu-repo/semantics/openAccessreponame:Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)instname:Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informaçãoinstacron:RCAAP2024-01-03T18:46:20Zoai:dspace.uevora.pt:10174/6790Portal AgregadorONGhttps://www.rcaap.pt/oai/openaireopendoar:71602024-03-20T01:01:25.752510Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) - Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informaçãofalse
dc.title.none.fl_str_mv Satellite DNA as a target for TaqMan real-time PCR detection of the pinewood nematode, Bursaphelenchus xylophilus
title Satellite DNA as a target for TaqMan real-time PCR detection of the pinewood nematode, Bursaphelenchus xylophilus
spellingShingle Satellite DNA as a target for TaqMan real-time PCR detection of the pinewood nematode, Bursaphelenchus xylophilus
CECILE, FRANÇOIS
Bursaphelenchus xylophilus
pinewood
nematode
title_short Satellite DNA as a target for TaqMan real-time PCR detection of the pinewood nematode, Bursaphelenchus xylophilus
title_full Satellite DNA as a target for TaqMan real-time PCR detection of the pinewood nematode, Bursaphelenchus xylophilus
title_fullStr Satellite DNA as a target for TaqMan real-time PCR detection of the pinewood nematode, Bursaphelenchus xylophilus
title_full_unstemmed Satellite DNA as a target for TaqMan real-time PCR detection of the pinewood nematode, Bursaphelenchus xylophilus
title_sort Satellite DNA as a target for TaqMan real-time PCR detection of the pinewood nematode, Bursaphelenchus xylophilus
author CECILE, FRANÇOIS
author_facet CECILE, FRANÇOIS
CASTAGNONE, CHANTAL
BOONHAM, NEIL
TOMLINSON, JENNY
LAWSON, REBECCA
HOCKLAND, SUE
QUILL, JAMES
VIEIRA, Paulo
Mota, Manuel
CASTAGNONE-SERENO, PHILIPPE
author_role author
author2 CASTAGNONE, CHANTAL
BOONHAM, NEIL
TOMLINSON, JENNY
LAWSON, REBECCA
HOCKLAND, SUE
QUILL, JAMES
VIEIRA, Paulo
Mota, Manuel
CASTAGNONE-SERENO, PHILIPPE
author2_role author
author
author
author
author
author
author
author
author
dc.contributor.author.fl_str_mv CECILE, FRANÇOIS
CASTAGNONE, CHANTAL
BOONHAM, NEIL
TOMLINSON, JENNY
LAWSON, REBECCA
HOCKLAND, SUE
QUILL, JAMES
VIEIRA, Paulo
Mota, Manuel
CASTAGNONE-SERENO, PHILIPPE
dc.subject.por.fl_str_mv Bursaphelenchus xylophilus
pinewood
nematode
topic Bursaphelenchus xylophilus
pinewood
nematode
description The pinewood nematode (PWN), Bursaphelenchus xylophilus , is a major pathogen of conifers, which impacts on forest health, natural ecosystem stability and international trade. As a consequence, it has been listed as a quarantine organism in Europe. A real-time PCR approach based on TaqMan chemistry was developed to detect this organism. Specific probe and primers were designed based on the sequence of the Msp I satellite DNA family previously characterized in the genome of the nematode. The method proved to be specific in tests with target DNA from PWN isolates from worldwide origin. From a practical point of view, detection limit was 1 pg of target DNA or one individual nematode. In addition, PWN genomic DNA or single individuals were positively detected in mixed samples in which B. xylophilius was associated with the closely related non-pathogenic species B. mucronatus , up to the limit of 0.01% or 1% of the mixture, respectively. The real-time PCR assay was also used in conjunction with a simple DNA extraction method to detect PWN directly in artificially infested wood samples. These results demonstrate the potential of this assay to provide rapid, accurate and sensitive molecular identification of the PWN in relation to pest risk assessment in the field and quarantine regulation.
publishDate 2007
dc.date.none.fl_str_mv 2007-01-01T00:00:00Z
2012-12-10T16:25:43Z
2012-12-10
dc.type.status.fl_str_mv info:eu-repo/semantics/publishedVersion
dc.type.driver.fl_str_mv info:eu-repo/semantics/article
format article
status_str publishedVersion
dc.identifier.uri.fl_str_mv http://hdl.handle.net/10174/6790
http://hdl.handle.net/10174/6790
url http://hdl.handle.net/10174/6790
dc.language.iso.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv 8
ICAAM
nd
nd
nd
nd
nd
nd
nd
pvieira@uevora.pt
mmtota@uevora.pt
Philippe.Castagnone@sophia.inra.fr
218
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