A new molecular method for the rapid subtyping of bovine herpesvirus 1 field isolates

Bovine herpesvirus 1 (BoHV-1) causes several clinical syndromes in cattle worldwide. There are 3 subtypes of BoHV-1: 1.1, 1.2a, and 1.2b. Several molecular methods are commonly used in the detection and characterization of BoHV-1. Among them, restriction endonuclease analysis (REA) and single-nucleo...

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Autores principales: Maidana, Silvina Soledad, Miño, Samuel, Apostolo, Romina Maria, De Stefano, Gabriel Alejandro, Romera, Sonia Alejandra
Formato: Artículo
Lenguaje:Inglés
Publicado: Sage Publications 2020
Materias:
Acceso en línea:http://hdl.handle.net/20.500.12123/7868
https://journals.sagepub.com/doi/abs/10.1177/1040638719898692
https://doi.org/10.1177/1040638719898692
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author Maidana, Silvina Soledad
Miño, Samuel
Apostolo, Romina Maria
De Stefano, Gabriel Alejandro
Romera, Sonia Alejandra
author_browse Apostolo, Romina Maria
De Stefano, Gabriel Alejandro
Maidana, Silvina Soledad
Miño, Samuel
Romera, Sonia Alejandra
author_facet Maidana, Silvina Soledad
Miño, Samuel
Apostolo, Romina Maria
De Stefano, Gabriel Alejandro
Romera, Sonia Alejandra
author_sort Maidana, Silvina Soledad
collection INTA Digital
description Bovine herpesvirus 1 (BoHV-1) causes several clinical syndromes in cattle worldwide. There are 3 subtypes of BoHV-1: 1.1, 1.2a, and 1.2b. Several molecular methods are commonly used in the detection and characterization of BoHV-1. Among them, restriction endonuclease analysis (REA) and single-nucleotide polymorphism (SNP) analysis of the complete viral genome allow classification of BoHV-1 into different subtypes. However, developing countries need simpler and cheaper screening assays for routine testing. We designed a standard multiplex PCR followed by a REA assay allowing straightforward subclassification of all BoHV-1 isolates tested into 1.1, 1.2a, and 1.2b subtypes based on the analysis of fragment length polymorphism. Our standard multiplex PCR-REA was used to analyze 33 field strains of BoHV-1 isolated from various tissues. The assay confirmed the subtype identified previously by REA. In addition, non-polymorphic or undigested fragments were sequenced in order to confirm the mutation affecting the RE HindIII site. Our PCR-REA method is an affordable and rapid test that will subtype all BoHV-1 strains.
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institution Instituto Nacional de Tecnología Agropecuaria (INTA -Argentina)
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spelling INTA78682020-09-10T16:53:09Z A new molecular method for the rapid subtyping of bovine herpesvirus 1 field isolates Maidana, Silvina Soledad Miño, Samuel Apostolo, Romina Maria De Stefano, Gabriel Alejandro Romera, Sonia Alejandra Bovine Herpesvirus Genetic Techniques Genetic Isolation Herpes Virus Bovino PCR Técnica Genética Aislamiento Genético BoHV-1 PCR Múltiple Nuevo Método Molecular Multiplex PCR New Molecular Method Bovine herpesvirus 1 (BoHV-1) causes several clinical syndromes in cattle worldwide. There are 3 subtypes of BoHV-1: 1.1, 1.2a, and 1.2b. Several molecular methods are commonly used in the detection and characterization of BoHV-1. Among them, restriction endonuclease analysis (REA) and single-nucleotide polymorphism (SNP) analysis of the complete viral genome allow classification of BoHV-1 into different subtypes. However, developing countries need simpler and cheaper screening assays for routine testing. We designed a standard multiplex PCR followed by a REA assay allowing straightforward subclassification of all BoHV-1 isolates tested into 1.1, 1.2a, and 1.2b subtypes based on the analysis of fragment length polymorphism. Our standard multiplex PCR-REA was used to analyze 33 field strains of BoHV-1 isolated from various tissues. The assay confirmed the subtype identified previously by REA. In addition, non-polymorphic or undigested fragments were sequenced in order to confirm the mutation affecting the RE HindIII site. Our PCR-REA method is an affordable and rapid test that will subtype all BoHV-1 strains. Instituto de Virología Fil: Maidana, Silvina Soledad. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Virología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Miño, Samuel. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Virología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Apostolo, Romina Maria. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Esquel. Grupo de Producción Animal; Argentina Fil: De Stefano, Gabriel Alejandro. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Virología; Argentina Fil: Romera, Sonia Alejandra. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Virología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad del Salvador. Cátedra de Inmunología; Argentina. Universidad de Morón. Facultad de Ciencias exactas, Químicas y Naturales. Cátedra de Inmunogenética; Argentina 2020-09-10T16:40:56Z 2020-09-10T16:40:56Z 2020-02 info:ar-repo/semantics/artículo info:eu-repo/semantics/article info:eu-repo/semantics/acceptedVersion http://hdl.handle.net/20.500.12123/7868 https://journals.sagepub.com/doi/abs/10.1177/1040638719898692 1943-4936 1040-6387 https://doi.org/10.1177/1040638719898692 eng info:eu-repograntAgreement/INTA/PNSA/1115054/AR./Enfermedades parasitarias, infecciosas y tóxico metabólicas que afectan la productividad de los bóvidos para producción de carne y leche. info:eu-repograntAgreement/INTA/PNSA/1115055/AR./Enfermedades infecciosas, parasitarias y toxico metabólicas que afectan la productividad de los ovinos, caprinos y camélidos. info:eu-repo/semantics/restrictedAccess application/pdf Sage Publications Journal of Veterinary Diagnostic Investigation 32 (1) : 112-117 (Febrero 2020)
spellingShingle Bovine Herpesvirus
Genetic Techniques
Genetic Isolation
Herpes Virus Bovino
PCR
Técnica Genética
Aislamiento Genético
BoHV-1
PCR Múltiple
Nuevo Método Molecular
Multiplex PCR
New Molecular Method
Maidana, Silvina Soledad
Miño, Samuel
Apostolo, Romina Maria
De Stefano, Gabriel Alejandro
Romera, Sonia Alejandra
A new molecular method for the rapid subtyping of bovine herpesvirus 1 field isolates
title A new molecular method for the rapid subtyping of bovine herpesvirus 1 field isolates
title_full A new molecular method for the rapid subtyping of bovine herpesvirus 1 field isolates
title_fullStr A new molecular method for the rapid subtyping of bovine herpesvirus 1 field isolates
title_full_unstemmed A new molecular method for the rapid subtyping of bovine herpesvirus 1 field isolates
title_short A new molecular method for the rapid subtyping of bovine herpesvirus 1 field isolates
title_sort new molecular method for the rapid subtyping of bovine herpesvirus 1 field isolates
topic Bovine Herpesvirus
Genetic Techniques
Genetic Isolation
Herpes Virus Bovino
PCR
Técnica Genética
Aislamiento Genético
BoHV-1
PCR Múltiple
Nuevo Método Molecular
Multiplex PCR
New Molecular Method
url http://hdl.handle.net/20.500.12123/7868
https://journals.sagepub.com/doi/abs/10.1177/1040638719898692
https://doi.org/10.1177/1040638719898692
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