Multiplex PCR followed by restriction length polymorphism for the subtyping of bovine herpesvirus 5 isolates

Autores
Maidana, Silvina Soledad; Morano, Cintia Débora; Cianfrini, Daniela; Campos, Fabrício Souza; Roehe, Paulo Michel; Siedler, Bianca; De Stefano, Gabriel; Mauroy, Axel; Thiry, Etienne; Romera, Sonia
Año de publicación
2013
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
Abstract Background: Several types and subtypes of bovine herpesviruses 1 and 5 (BoHV-1 and BoHV-5) have been associated to different clinical conditions of cattle, making type/subtype differentiation essential to understand the pathogenesis and epidemiology of BoHV infections. BoHV-5 subtyping is currently carried out by BstEII restriction enzyme analysis (REA) of the complete virus genome. This method allowed the description of three subtypes, one of which is the most widespread while the remaining two have so far only been found in South America. The present work describes a multiplex PCR followed by REA for BoHV-5 subtyping. Results: The method consists in the simultaneous amplification of glycoprotein B and UL54 gene fragments of 534 and 669 base pairs (bp), respectively, BstEII digestion of amplicons, separation of products in 1% agarose gels, and analysis of fragment length polymorphims. The multiplex PCR detected up to 227 BoHV-5 genome copies and 9.2 × 105 BoHV-5 genome copies when DNA was extracted from purified virus or infected tissue homogenates, respectively. The applicability of multiplex PCR-REA was demonstrated on 3 BoHV-5 reference strains. In addition, subtyping of two new isolates and seventeen previously reported ones (17 BHV-5a and 2 BHV-5b) by this method gave coincident results with those obtained with the classic BstEII REA assay. Conclusions: Multiplex PCR-REA provides a new tool for the fast and simple diagnosis and subtyping of BoHV-5.
Fil: Maidana, Silvina Soledad. Instituto Nacional de Tecnología Agropecuaria. Centro Nacional de Investigaciones Agropecuarias. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Virología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Morano, Cintia Débora. Instituto Nacional de Tecnología Agropecuaria. Centro Nacional de Investigaciones Agropecuarias. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Virología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Cianfrini, Daniela. Tecnovax SA; Argentina
Fil: Campos, Fabrício Souza. Universidade Federal do Rio Grande do Sul; Brasil
Fil: Roehe, Paulo Michel. Universidade Federal do Rio Grande do Sul; Brasil
Fil: Siedler, Bianca. Universidade Federal de Pelotas; Brasil
Fil: De Stefano, Gabriel. Instituto Nacional de Tecnología Agropecuaria. Centro Nacional de Investigaciones Agropecuarias. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Virología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Mauroy, Axel. Universite de Liege; Bélgica
Fil: Thiry, Etienne. Universite de Liege; Bélgica
Fil: Romera, Sonia. Instituto Nacional de Tecnología Agropecuaria. Centro Nacional de Investigaciones Agropecuarias. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Virología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad del Salvador; Argentina
Materia
PCR
Multiplex
BoHV5
Nivel de accesibilidad
acceso abierto
Condiciones de uso
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
Repositorio
CONICET Digital (CONICET)
Institución
Consejo Nacional de Investigaciones Científicas y Técnicas
OAI Identificador
oai:ri.conicet.gov.ar:11336/11024
Acceder
id CONICETDig_282d17f2f02930e4798faff3c094f320
oai_identifier_str oai:ri.conicet.gov.ar:11336/11024
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling Multiplex PCR followed by restriction length polymorphism for the subtyping of bovine herpesvirus 5 isolatesMaidana, Silvina SoledadMorano, Cintia DéboraCianfrini, DanielaCampos, Fabrício SouzaRoehe, Paulo MichelSiedler, BiancaDe Stefano, GabrielMauroy, AxelThiry, EtienneRomera, SoniaPCRMultiplexBoHV5https://purl.org/becyt/ford/1.6https://purl.org/becyt/ford/1Abstract Background: Several types and subtypes of bovine herpesviruses 1 and 5 (BoHV-1 and BoHV-5) have been associated to different clinical conditions of cattle, making type/subtype differentiation essential to understand the pathogenesis and epidemiology of BoHV infections. BoHV-5 subtyping is currently carried out by BstEII restriction enzyme analysis (REA) of the complete virus genome. This method allowed the description of three subtypes, one of which is the most widespread while the remaining two have so far only been found in South America. The present work describes a multiplex PCR followed by REA for BoHV-5 subtyping. Results: The method consists in the simultaneous amplification of glycoprotein B and UL54 gene fragments of 534 and 669 base pairs (bp), respectively, BstEII digestion of amplicons, separation of products in 1% agarose gels, and analysis of fragment length polymorphims. The multiplex PCR detected up to 227 BoHV-5 genome copies and 9.2 × 105 BoHV-5 genome copies when DNA was extracted from purified virus or infected tissue homogenates, respectively. The applicability of multiplex PCR-REA was demonstrated on 3 BoHV-5 reference strains. In addition, subtyping of two new isolates and seventeen previously reported ones (17 BHV-5a and 2 BHV-5b) by this method gave coincident results with those obtained with the classic BstEII REA assay. Conclusions: Multiplex PCR-REA provides a new tool for the fast and simple diagnosis and subtyping of BoHV-5.Fil: Maidana, Silvina Soledad. Instituto Nacional de Tecnología Agropecuaria. Centro Nacional de Investigaciones Agropecuarias. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Virología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Morano, Cintia Débora. Instituto Nacional de Tecnología Agropecuaria. Centro Nacional de Investigaciones Agropecuarias. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Virología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Cianfrini, Daniela. Tecnovax SA; ArgentinaFil: Campos, Fabrício Souza. Universidade Federal do Rio Grande do Sul; BrasilFil: Roehe, Paulo Michel. Universidade Federal do Rio Grande do Sul; BrasilFil: Siedler, Bianca. Universidade Federal de Pelotas; BrasilFil: De Stefano, Gabriel. Instituto Nacional de Tecnología Agropecuaria. Centro Nacional de Investigaciones Agropecuarias. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Virología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Mauroy, Axel. Universite de Liege; BélgicaFil: Thiry, Etienne. Universite de Liege; BélgicaFil: Romera, Sonia. Instituto Nacional de Tecnología Agropecuaria. Centro Nacional de Investigaciones Agropecuarias. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Virología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad del Salvador; ArgentinaBiomed Central2013-06info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/11024Maidana, Silvina Soledad; Morano, Cintia Débora; Cianfrini, Daniela; Campos, Fabrício Souza; Roehe, Paulo Michel; et al.; Multiplex PCR followed by restriction length polymorphism for the subtyping of bovine herpesvirus 5 isolates; Biomed Central; Bmc Veterinary Research; 9; 6-2013; 111-1161746-6148enginfo:eu-repo/semantics/altIdentifier/url/https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3679755/info:eu-repo/semantics/altIdentifier/doi/10.1186/1746-6148-9-111info:eu-repo/semantics/altIdentifier/url/http://bmcvetres.biomedcentral.com/articles/10.1186/1746-6148-9-111info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-09-29T10:25:21Zoai:ri.conicet.gov.ar:11336/11024instacron: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 10:25:21.776CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Multiplex PCR followed by restriction length polymorphism for the subtyping of bovine herpesvirus 5 isolates
title Multiplex PCR followed by restriction length polymorphism for the subtyping of bovine herpesvirus 5 isolates
spellingShingle Multiplex PCR followed by restriction length polymorphism for the subtyping of bovine herpesvirus 5 isolates
Maidana, Silvina Soledad
PCR
Multiplex
BoHV5
title_short Multiplex PCR followed by restriction length polymorphism for the subtyping of bovine herpesvirus 5 isolates
title_full Multiplex PCR followed by restriction length polymorphism for the subtyping of bovine herpesvirus 5 isolates
title_fullStr Multiplex PCR followed by restriction length polymorphism for the subtyping of bovine herpesvirus 5 isolates
title_full_unstemmed Multiplex PCR followed by restriction length polymorphism for the subtyping of bovine herpesvirus 5 isolates
title_sort Multiplex PCR followed by restriction length polymorphism for the subtyping of bovine herpesvirus 5 isolates
dc.creator.none.fl_str_mv Maidana, Silvina Soledad
Morano, Cintia Débora
Cianfrini, Daniela
Campos, Fabrício Souza
Roehe, Paulo Michel
Siedler, Bianca
De Stefano, Gabriel
Mauroy, Axel
Thiry, Etienne
Romera, Sonia
author Maidana, Silvina Soledad
author_facet Maidana, Silvina Soledad
Morano, Cintia Débora
Cianfrini, Daniela
Campos, Fabrício Souza
Roehe, Paulo Michel
Siedler, Bianca
De Stefano, Gabriel
Mauroy, Axel
Thiry, Etienne
Romera, Sonia
author_role author
author2 Morano, Cintia Débora
Cianfrini, Daniela
Campos, Fabrício Souza
Roehe, Paulo Michel
Siedler, Bianca
De Stefano, Gabriel
Mauroy, Axel
Thiry, Etienne
Romera, Sonia
author2_role author
author
author
author
author
author
author
author
author
dc.subject.none.fl_str_mv PCR
Multiplex
BoHV5
topic PCR
Multiplex
BoHV5
purl_subject.fl_str_mv https://purl.org/becyt/ford/1.6
https://purl.org/becyt/ford/1
dc.description.none.fl_txt_mv Abstract Background: Several types and subtypes of bovine herpesviruses 1 and 5 (BoHV-1 and BoHV-5) have been associated to different clinical conditions of cattle, making type/subtype differentiation essential to understand the pathogenesis and epidemiology of BoHV infections. BoHV-5 subtyping is currently carried out by BstEII restriction enzyme analysis (REA) of the complete virus genome. This method allowed the description of three subtypes, one of which is the most widespread while the remaining two have so far only been found in South America. The present work describes a multiplex PCR followed by REA for BoHV-5 subtyping. Results: The method consists in the simultaneous amplification of glycoprotein B and UL54 gene fragments of 534 and 669 base pairs (bp), respectively, BstEII digestion of amplicons, separation of products in 1% agarose gels, and analysis of fragment length polymorphims. The multiplex PCR detected up to 227 BoHV-5 genome copies and 9.2 × 105 BoHV-5 genome copies when DNA was extracted from purified virus or infected tissue homogenates, respectively. The applicability of multiplex PCR-REA was demonstrated on 3 BoHV-5 reference strains. In addition, subtyping of two new isolates and seventeen previously reported ones (17 BHV-5a and 2 BHV-5b) by this method gave coincident results with those obtained with the classic BstEII REA assay. Conclusions: Multiplex PCR-REA provides a new tool for the fast and simple diagnosis and subtyping of BoHV-5.
Fil: Maidana, Silvina Soledad. Instituto Nacional de Tecnología Agropecuaria. Centro Nacional de Investigaciones Agropecuarias. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Virología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Morano, Cintia Débora. Instituto Nacional de Tecnología Agropecuaria. Centro Nacional de Investigaciones Agropecuarias. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Virología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Cianfrini, Daniela. Tecnovax SA; Argentina
Fil: Campos, Fabrício Souza. Universidade Federal do Rio Grande do Sul; Brasil
Fil: Roehe, Paulo Michel. Universidade Federal do Rio Grande do Sul; Brasil
Fil: Siedler, Bianca. Universidade Federal de Pelotas; Brasil
Fil: De Stefano, Gabriel. Instituto Nacional de Tecnología Agropecuaria. Centro Nacional de Investigaciones Agropecuarias. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Virología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Mauroy, Axel. Universite de Liege; Bélgica
Fil: Thiry, Etienne. Universite de Liege; Bélgica
Fil: Romera, Sonia. Instituto Nacional de Tecnología Agropecuaria. Centro Nacional de Investigaciones Agropecuarias. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Virología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad del Salvador; Argentina
description Abstract Background: Several types and subtypes of bovine herpesviruses 1 and 5 (BoHV-1 and BoHV-5) have been associated to different clinical conditions of cattle, making type/subtype differentiation essential to understand the pathogenesis and epidemiology of BoHV infections. BoHV-5 subtyping is currently carried out by BstEII restriction enzyme analysis (REA) of the complete virus genome. This method allowed the description of three subtypes, one of which is the most widespread while the remaining two have so far only been found in South America. The present work describes a multiplex PCR followed by REA for BoHV-5 subtyping. Results: The method consists in the simultaneous amplification of glycoprotein B and UL54 gene fragments of 534 and 669 base pairs (bp), respectively, BstEII digestion of amplicons, separation of products in 1% agarose gels, and analysis of fragment length polymorphims. The multiplex PCR detected up to 227 BoHV-5 genome copies and 9.2 × 105 BoHV-5 genome copies when DNA was extracted from purified virus or infected tissue homogenates, respectively. The applicability of multiplex PCR-REA was demonstrated on 3 BoHV-5 reference strains. In addition, subtyping of two new isolates and seventeen previously reported ones (17 BHV-5a and 2 BHV-5b) by this method gave coincident results with those obtained with the classic BstEII REA assay. Conclusions: Multiplex PCR-REA provides a new tool for the fast and simple diagnosis and subtyping of BoHV-5.
publishDate 2013
dc.date.none.fl_str_mv 2013-06
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/11336/11024
Maidana, Silvina Soledad; Morano, Cintia Débora; Cianfrini, Daniela; Campos, Fabrício Souza; Roehe, Paulo Michel; et al.; Multiplex PCR followed by restriction length polymorphism for the subtyping of bovine herpesvirus 5 isolates; Biomed Central; Bmc Veterinary Research; 9; 6-2013; 111-116
1746-6148
url http://hdl.handle.net/11336/11024
identifier_str_mv Maidana, Silvina Soledad; Morano, Cintia Débora; Cianfrini, Daniela; Campos, Fabrício Souza; Roehe, Paulo Michel; et al.; Multiplex PCR followed by restriction length polymorphism for the subtyping of bovine herpesvirus 5 isolates; Biomed Central; Bmc Veterinary Research; 9; 6-2013; 111-116
1746-6148
dc.language.none.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv info:eu-repo/semantics/altIdentifier/url/https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3679755/
info:eu-repo/semantics/altIdentifier/doi/10.1186/1746-6148-9-111
info:eu-repo/semantics/altIdentifier/url/http://bmcvetres.biomedcentral.com/articles/10.1186/1746-6148-9-111
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
eu_rights_str_mv openAccess
rights_invalid_str_mv https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
dc.format.none.fl_str_mv application/pdf
application/pdf
dc.publisher.none.fl_str_mv Biomed Central
publisher.none.fl_str_mv Biomed Central
dc.source.none.fl_str_mv reponame:CONICET Digital (CONICET)
instname:Consejo Nacional de Investigaciones Científicas y Técnicas
reponame_str CONICET Digital (CONICET)
collection CONICET Digital (CONICET)
instname_str Consejo Nacional de Investigaciones Científicas y Técnicas
repository.name.fl_str_mv CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicas
repository.mail.fl_str_mv dasensio@conicet.gov.ar; lcarlino@conicet.gov.ar
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score 13.070432

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