A 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) analysis

Autores
Marin, Maia Solange; Quintana, Silvina; Leunda, Maria Rosa; Recavarren, Mariana Ines; Pagnuco, Inti Anabela; Spath, Ernesto Juan; Perez, Sandra; Odeon, Anselmo Carlos
Año de publicación
2016
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
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 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.
EEA Balcarce
Fil: Marin, Maia Solange. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Quintana, Silvina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Instituto de Análisis Fares Taie; Argentina
Fil: Leunda, Maria Rosa. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Balcarce; Argentina
Fil: Recavarren, Mariana Ines. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Instituto de Análisis Fares Taie; Argentina
Fil: Pagnuco, Inti Anabela. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Spath, Ernesto Juan. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Balcarce; Argentina
Fil: 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; Argentina
Fil: Odeón, Anselmo Carlos. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Balcarce; Argentina
Fuente
Journal of Virological Methods 227 : 14-22 (January 2016)
Materia
Ganado Bovino
Enfermedades de los Animales
Herpes Virus Bovino
PCR
Diagnóstico
Cattle
Animal Diseases
Bovine Herpesvirus
Diagnosis
Nivel de accesibilidad
acceso restringido
Condiciones de uso
Repositorio
INTA Digital (INTA)
Institución
Instituto Nacional de Tecnología Agropecuaria
OAI Identificador
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oai_identifier_str oai:localhost:20.500.12123/4981
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spelling A 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) analysisMarin, Maia SolangeQuintana, SilvinaLeunda, Maria RosaRecavarren, Mariana InesPagnuco, Inti AnabelaSpath, Ernesto JuanPerez, SandraOdeon, Anselmo CarlosGanado BovinoEnfermedades de los AnimalesHerpes Virus BovinoPCRDiagnósticoCattleAnimal DiseasesBovine HerpesvirusDiagnosisBovine 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.EEA BalcarceFil: Marin, Maia Solange. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Quintana, Silvina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Instituto de Análisis Fares Taie; ArgentinaFil: Leunda, Maria Rosa. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Balcarce; ArgentinaFil: Recavarren, Mariana Ines. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Instituto de Análisis Fares Taie; ArgentinaFil: Pagnuco, Inti Anabela. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Spath, Ernesto Juan. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Balcarce; ArgentinaFil: 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; ArgentinaFil: Odeón, Anselmo Carlos. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Balcarce; ArgentinaElsevier2019-04-25T13:08:56Z2019-04-25T13:08:56Z2016-01info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfhttps://www.sciencedirect.com/science/article/pii/S0166093415003390http://hdl.handle.net/20.500.12123/49810166-0934https://doi.org/10.1016/j.jviromet.2015.10.005Journal of Virological Methods 227 : 14-22 (January 2016)reponame:INTA Digital (INTA)instname:Instituto Nacional de Tecnología Agropecuariaenginfo:eu-repo/semantics/restrictedAccess2025-09-04T09:47:56Zoai:localhost:20.500.12123/4981instacron:INTAInstitucionalhttp://repositorio.inta.gob.ar/Organismo científico-tecnológicoNo correspondehttp://repositorio.inta.gob.ar/oai/requesttripaldi.nicolas@inta.gob.arArgentinaNo correspondeNo correspondeNo correspondeopendoar:l2025-09-04 09:47:57.298INTA Digital (INTA) - Instituto Nacional de Tecnología Agropecuariafalse
dc.title.none.fl_str_mv A 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) analysis
title A 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) analysis
spellingShingle A 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) analysis
Marin, Maia Solange
Ganado Bovino
Enfermedades de los Animales
Herpes Virus Bovino
PCR
Diagnóstico
Cattle
Animal Diseases
Bovine Herpesvirus
Diagnosis
title_short A 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) analysis
title_full A 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) analysis
title_fullStr A 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) analysis
title_full_unstemmed A 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) analysis
title_sort A 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) analysis
dc.creator.none.fl_str_mv Marin, Maia Solange
Quintana, Silvina
Leunda, Maria Rosa
Recavarren, Mariana Ines
Pagnuco, Inti Anabela
Spath, Ernesto Juan
Perez, Sandra
Odeon, Anselmo Carlos
author Marin, Maia Solange
author_facet Marin, Maia Solange
Quintana, Silvina
Leunda, Maria Rosa
Recavarren, Mariana Ines
Pagnuco, Inti Anabela
Spath, Ernesto Juan
Perez, Sandra
Odeon, Anselmo Carlos
author_role author
author2 Quintana, Silvina
Leunda, Maria Rosa
Recavarren, Mariana Ines
Pagnuco, Inti Anabela
Spath, Ernesto Juan
Perez, Sandra
Odeon, Anselmo Carlos
author2_role author
author
author
author
author
author
author
dc.subject.none.fl_str_mv Ganado Bovino
Enfermedades de los Animales
Herpes Virus Bovino
PCR
Diagnóstico
Cattle
Animal Diseases
Bovine Herpesvirus
Diagnosis
topic Ganado Bovino
Enfermedades de los Animales
Herpes Virus Bovino
PCR
Diagnóstico
Cattle
Animal Diseases
Bovine Herpesvirus
Diagnosis
dc.description.none.fl_txt_mv 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 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.
EEA Balcarce
Fil: Marin, Maia Solange. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Quintana, Silvina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Instituto de Análisis Fares Taie; Argentina
Fil: Leunda, Maria Rosa. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Balcarce; Argentina
Fil: Recavarren, Mariana Ines. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Instituto de Análisis Fares Taie; Argentina
Fil: Pagnuco, Inti Anabela. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Spath, Ernesto Juan. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Balcarce; Argentina
Fil: 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; Argentina
Fil: Odeón, Anselmo Carlos. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Balcarce; Argentina
description 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 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.
publishDate 2016
dc.date.none.fl_str_mv 2016-01
2019-04-25T13:08:56Z
2019-04-25T13:08:56Z
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 https://www.sciencedirect.com/science/article/pii/S0166093415003390
http://hdl.handle.net/20.500.12123/4981
0166-0934
https://doi.org/10.1016/j.jviromet.2015.10.005
url https://www.sciencedirect.com/science/article/pii/S0166093415003390
http://hdl.handle.net/20.500.12123/4981
https://doi.org/10.1016/j.jviromet.2015.10.005
identifier_str_mv 0166-0934
dc.language.none.fl_str_mv eng
language eng
dc.rights.none.fl_str_mv info:eu-repo/semantics/restrictedAccess
eu_rights_str_mv restrictedAccess
dc.format.none.fl_str_mv application/pdf
dc.publisher.none.fl_str_mv Elsevier
publisher.none.fl_str_mv Elsevier
dc.source.none.fl_str_mv Journal of Virological Methods 227 : 14-22 (January 2016)
reponame:INTA Digital (INTA)
instname:Instituto Nacional de Tecnología Agropecuaria
reponame_str INTA Digital (INTA)
collection INTA Digital (INTA)
instname_str Instituto Nacional de Tecnología Agropecuaria
repository.name.fl_str_mv INTA Digital (INTA) - Instituto Nacional de Tecnología Agropecuaria
repository.mail.fl_str_mv tripaldi.nicolas@inta.gob.ar
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score 12.623145

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