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dc.contributor.author
Maidana, Silvina Soledad
dc.contributor.author
Morano, Cintia Débora
dc.contributor.author
Cianfrini, Daniela
dc.contributor.author
Campos, Fabrício Souza
dc.contributor.author
Roehe, Paulo Michel
dc.contributor.author
Siedler, Bianca
dc.contributor.author
De Stefano, Gabriel
dc.contributor.author
Mauroy, Axel
dc.contributor.author
Thiry, Etienne
dc.contributor.author
Romera, Sonia
dc.date.available
2017-01-10T14:34:17Z
dc.date.issued
2013-06
dc.identifier.citation
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
dc.identifier.issn
1746-6148
dc.identifier.uri
http://hdl.handle.net/11336/11024
dc.description.abstract
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.
dc.format
application/pdf
dc.language.iso
eng
dc.publisher
Biomed Central
dc.rights
info:eu-repo/semantics/openAccess
dc.rights.uri
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
dc.subject
Pcr
dc.subject
Multiplex
dc.subject
Bohv5
dc.subject.classification
Virología
dc.subject.classification
Ciencias Biológicas
dc.subject.classification
CIENCIAS NATURALES Y EXACTAS
dc.title
Multiplex PCR followed by restriction length polymorphism for the subtyping of bovine herpesvirus 5 isolates
dc.type
info:eu-repo/semantics/article
dc.type
info:ar-repo/semantics/artículo
dc.type
info:eu-repo/semantics/publishedVersion
dc.date.updated
2016-11-24T18:38:54Z
dc.journal.volume
9
dc.journal.pagination
111-116
dc.journal.pais
Reino Unido
dc.journal.ciudad
Londres
dc.description.fil
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
dc.description.fil
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
dc.description.fil
Fil: Cianfrini, Daniela. Tecnovax SA; Argentina
dc.description.fil
Fil: Campos, Fabrício Souza. Universidade Federal do Rio Grande do Sul; Brasil
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Fil: Roehe, Paulo Michel. Universidade Federal do Rio Grande do Sul; Brasil
dc.description.fil
Fil: Siedler, Bianca. Universidade Federal de Pelotas; Brasil
dc.description.fil
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
dc.description.fil
Fil: Mauroy, Axel. Universite de Liege; Bélgica
dc.description.fil
Fil: Thiry, Etienne. Universite de Liege; Bélgica
dc.description.fil
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
dc.journal.title
Bmc Veterinary Research
dc.relation.alternativeid
info:eu-repo/semantics/altIdentifier/url/https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3679755/
dc.relation.alternativeid
info:eu-repo/semantics/altIdentifier/doi/http://dx.doi.org/10.1186/1746-6148-9-111
dc.relation.alternativeid
info:eu-repo/semantics/altIdentifier/url/http://bmcvetres.biomedcentral.com/articles/10.1186/1746-6148-9-111


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