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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
dc.description.fil 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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info:eu-repo/semantics/openAccess Excepto donde se diga explícitamente, este item se publica bajo la siguiente descripción: Creative Commons Attribution-NonCommercial-ShareAlike 2.5 Unported (CC BY-NC-SA 2.5)