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Abstract
Bovine herpesvirus types 1 (BoHV-1) and 5 (BoHV-5) are antigenically and genetically similar. The aim of this study was to develop a simple and reliable one-step real time PCR assay with high resolution melting (HRM) analysis for the simultaneous detection and differentiation of BoHV-1 and BoHV-5. Optimization of assay conditions was performed with DNA from reference strains. Then, DNA from field isolates, clinical samples and tissue samples of [ver mas...]
dc.contributor.authorMarin, Maia Solange
dc.contributor.authorQuintana, Silvina
dc.contributor.authorLeunda, Maria Rosa
dc.contributor.authorRecavarren, Mariana Ines
dc.contributor.authorPagnuco, Inti Anabela
dc.contributor.authorSpath, Ernesto Juan
dc.contributor.authorPerez, Sandra
dc.contributor.authorOdeon, Anselmo Carlos
dc.date.accessioned2019-04-25T13:08:56Z
dc.date.available2019-04-25T13:08:56Z
dc.date.issued2016-01
dc.identifier.issn0166-0934
dc.identifier.otherhttps://doi.org/10.1016/j.jviromet.2015.10.005
dc.identifier.urihttps://www.sciencedirect.com/science/article/pii/S0166093415003390
dc.identifier.urihttp://hdl.handle.net/20.500.12123/4981
dc.description.abstractBovine herpesvirus types 1 (BoHV-1) and 5 (BoHV-5) are antigenically and genetically similar. The aim of this study was to develop a simple and reliable one-step real time PCR assay with high resolution melting (HRM) analysis for the simultaneous detection and differentiation of BoHV-1 and BoHV-5. Optimization of assay conditions was performed with DNA from reference strains. Then, DNA from field isolates, clinical samples and tissue samples of experimentally infected animals were studied by real time PCR-HRM. An efficient amplification of real time PCR products was obtained, and a clear melting curve and appropriate melting peaks for both viruses were achieved in the HRM curve analysis for BoHV type identification. BoHV was identified in all of the isolates and clinical samples, and BoHV types were properly differentiated. Furthermore, viral DNA was detected in 12/18 and 7/18 samples from BoHV-1- and BoHV-5-infected calves, respectively. Real time PCR-HRM achieved a higher sensitivity compared with virus isolation or conventional PCR. In this study, HRM was used as a novel procedure. This method provides rapid, sensitive, specific and simultaneous detection of bovine alpha-herpesviruses DNA. Thus, this technique is an excellent tool for diagnosis, research and epidemiological studies of these viruses in cattle.eng
dc.formatapplication/pdfes_AR
dc.language.isoenges_AR
dc.publisherElsevieres_AR
dc.rightsinfo:eu-repo/semantics/restrictedAccesses_AR
dc.sourceJournal of Virological Methods 227 : 14-22 (January 2016)es_AR
dc.subjectGanado Bovinoes_AR
dc.subjectCattleeng
dc.subjectEnfermedades de los Animaleses_AR
dc.subjectAnimal Diseaseseng
dc.subjectHerpes Virus Bovinoes_AR
dc.subjectBovine Herpesviruseng
dc.subjectPCRes_AR
dc.subjectDiagnósticoes_AR
dc.subjectDiagnosiseng
dc.titleA new method for simultaneous detection and discrimination of Bovine herpesvirus types 1 (BoHV-1) and 5 (BoHV-5) using real time PCR with high resolution melting (HRM) analysises_AR
dc.typeinfo:ar-repo/semantics/artículoes_AR
dc.typeinfo:eu-repo/semantics/articlees_AR
dc.typeinfo:eu-repo/semantics/publishedVersiones_AR
dc.description.origenEEA Balcarcees_AR
dc.description.filFil: Marin, Maia Solange. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentinaes_AR
dc.description.filFil: Quintana, Silvina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Instituto de Análisis Fares Taie; Argentinaes_AR
dc.description.filFil: Leunda, Maria Rosa. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Balcarce; Argentinaes_AR
dc.description.filFil: Recavarren, Mariana Ines. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Instituto de Análisis Fares Taie; Argentinaes_AR
dc.description.filFil: Pagnuco, Inti Anabela. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentinaes_AR
dc.description.filFil: Spath, Ernesto Juan. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Balcarce; Argentinaes_AR
dc.description.filFil: Perez, Sandra. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tandil. Centro de Investigación Veterinaria de Tandil. Universidad Nacional del Centro de la Provincia de Buenos Aires. Centro de Investigación Veterinaria de Tandil. Provincia de Buenos Aires. Gobernación. Comision de Investigaciones Científicas. Centro de Investigación Veterinaria de Tandil; Argentinaes_AR
dc.description.filFil: Odeón, Anselmo Carlos. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Balcarce; Argentinaes_AR
dc.subtypecientifico


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