Desarrollo y análisis del sistema conjunto pcr/dot blot para la detección del virus de la necrosis hipodérmica y hematopoyética infecciosa

The present work describes development and analysis of a PCR/dot blot system for the detection of IHHNV. The experimental strategy was performed in three phases: 1) cloning of specific regions of the viral genome for generation of oligonucleotide probes to recognize a region of the IHHNV genome, 2)...

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Detalhes bibliográficos
Autores: Castro Lara, Jonathan, Robalino Iturralde, Javier
Formato: artículo
Estado:Versión publicada
Fecha de publicación:2009
País:Ecuador
Recursos:Escuela Superior Politécnica del Litoral
Repositorio:Repositorio Escuela Superior Politécnica del Litoral
Idioma:español
OAI Identifier:oai:www.dspace.espol.edu.ec:123456789/6943
Acesso em linha:http://www.dspace.espol.edu.ec/handle/123456789/6943
Access Level:acceso abierto
Palavra-chave:IHHNV
DOT BLOT
PCR
SONDAS NUCLEICAS
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spelling Desarrollo y análisis del sistema conjunto pcr/dot blot para la detección del virus de la necrosis hipodérmica y hematopoyética infecciosaCastro Lara, JonathanRobalino Iturralde, JavierIHHNVDOT BLOTPCRSONDAS NUCLEICASThe present work describes development and analysis of a PCR/dot blot system for the detection of IHHNV. The experimental strategy was performed in three phases: 1) cloning of specific regions of the viral genome for generation of oligonucleotide probes to recognize a region of the IHHNV genome, 2) optimization of the PCR/dot blot protocol as a diagnostic tool, and 3) to test the method for diagnostic of IHHNV in Penaeus vannamei. The analysis of sensibility performed in fragments of purified viral DNA showed levels of dot blot detection (without PCR) of 3pg/μl. The use of PCR/dot blot system increased the sensibility of detection to 0.002ag/μl of target DNA, in comparison; the sensibility of PCR without dot blot was 2ag/μl. This increase in sensibility was evident in the laboratory test of P. vannamei samples. Broodstok population was analyzed using PCR and PCR/dot blot, these analyses showed positive signals for IHHNV in 69.83% and 79.9%, respectively. In a similar fashion, a population of juvenile shrimps showed positive signals for IHHNV in 7.48% using PCR and 13.3% using PCR/dot blot. These results support the use of PCR/dot blot system as an alternative method for the sensitive and specific detection of viral genome, as compared to the traditional PCR method.2009-08-252009-08-252009-08-25info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articleapplication/pdfhttp://www.dspace.espol.edu.ec/handle/123456789/6943spainfo:eu-repo/semantics/openAccessreponame:Repositorio Escuela Superior Politécnica del Litoralinstname:Escuela Superior Politécnica del Litoralinstacron:ESPOL2018-04-04T17:16:41Zoai:www.dspace.espol.edu.ec:123456789/6943Institucionalhttps://www.dspace.espol.edu.ec/Universidad públicahttps://www.espol.edu.ec/.https://www.dspace.espol.edu.ec/oaiEcuador...opendoar:14792024-07-27T05:53:21.360461trueInstitucionalhttps://www.dspace.espol.edu.ec/Universidad públicahttps://www.espol.edu.ec/.https://www.dspace.espol.edu.ec/oai.Ecuador...opendoar:14792024-07-27T05:53:21.360461Repositorio Escuela Superior Politécnica del Litoral - Escuela Superior Politécnica del Litoraltrue
dc.title.none.fl_str_mv Desarrollo y análisis del sistema conjunto pcr/dot blot para la detección del virus de la necrosis hipodérmica y hematopoyética infecciosa
title Desarrollo y análisis del sistema conjunto pcr/dot blot para la detección del virus de la necrosis hipodérmica y hematopoyética infecciosa
spellingShingle Desarrollo y análisis del sistema conjunto pcr/dot blot para la detección del virus de la necrosis hipodérmica y hematopoyética infecciosa
Castro Lara, Jonathan
IHHNV
DOT BLOT
PCR
SONDAS NUCLEICAS
title_short Desarrollo y análisis del sistema conjunto pcr/dot blot para la detección del virus de la necrosis hipodérmica y hematopoyética infecciosa
title_full Desarrollo y análisis del sistema conjunto pcr/dot blot para la detección del virus de la necrosis hipodérmica y hematopoyética infecciosa
title_fullStr Desarrollo y análisis del sistema conjunto pcr/dot blot para la detección del virus de la necrosis hipodérmica y hematopoyética infecciosa
title_full_unstemmed Desarrollo y análisis del sistema conjunto pcr/dot blot para la detección del virus de la necrosis hipodérmica y hematopoyética infecciosa
title_sort Desarrollo y análisis del sistema conjunto pcr/dot blot para la detección del virus de la necrosis hipodérmica y hematopoyética infecciosa
dc.creator.none.fl_str_mv Castro Lara, Jonathan
Robalino Iturralde, Javier
author Castro Lara, Jonathan
author_facet Castro Lara, Jonathan
Robalino Iturralde, Javier
author_role author
author2 Robalino Iturralde, Javier
author2_role author
dc.subject.none.fl_str_mv IHHNV
DOT BLOT
PCR
SONDAS NUCLEICAS
topic IHHNV
DOT BLOT
PCR
SONDAS NUCLEICAS
description The present work describes development and analysis of a PCR/dot blot system for the detection of IHHNV. The experimental strategy was performed in three phases: 1) cloning of specific regions of the viral genome for generation of oligonucleotide probes to recognize a region of the IHHNV genome, 2) optimization of the PCR/dot blot protocol as a diagnostic tool, and 3) to test the method for diagnostic of IHHNV in Penaeus vannamei. The analysis of sensibility performed in fragments of purified viral DNA showed levels of dot blot detection (without PCR) of 3pg/μl. The use of PCR/dot blot system increased the sensibility of detection to 0.002ag/μl of target DNA, in comparison; the sensibility of PCR without dot blot was 2ag/μl. This increase in sensibility was evident in the laboratory test of P. vannamei samples. Broodstok population was analyzed using PCR and PCR/dot blot, these analyses showed positive signals for IHHNV in 69.83% and 79.9%, respectively. In a similar fashion, a population of juvenile shrimps showed positive signals for IHHNV in 7.48% using PCR and 13.3% using PCR/dot blot. These results support the use of PCR/dot blot system as an alternative method for the sensitive and specific detection of viral genome, as compared to the traditional PCR method.
publishDate 2009
dc.date.none.fl_str_mv 2009-08-25
2009-08-25
2009-08-25
dc.type.none.fl_str_mv info:eu-repo/semantics/publishedVersion
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format article
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language spa
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
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dc.source.none.fl_str_mv reponame:Repositorio Escuela Superior Politécnica del Litoral
instname:Escuela Superior Politécnica del Litoral
instacron:ESPOL
instname_str Escuela Superior Politécnica del Litoral
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institution ESPOL
reponame_str Repositorio Escuela Superior Politécnica del Litoral
collection Repositorio Escuela Superior Politécnica del Litoral
repository.name.fl_str_mv Repositorio Escuela Superior Politécnica del Litoral - Escuela Superior Politécnica del Litoral
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