1. A new molecular method for the rapid subtyping of bovine herpesvirus 1 field isolates.
- Author
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Maidana SS, Miño S, Apostolo RM, De Stefano GA, and Romera SA
- Subjects
- Animals, Cattle, Cattle Diseases diagnosis, DNA, Viral analysis, Genome, Viral, Herpesviridae Infections diagnosis, Herpesvirus 1, Bovine genetics, Herpesvirus 1, Bovine isolation & purification, Multiplex Polymerase Chain Reaction veterinary, Mutation, Restriction Mapping, Cattle Diseases virology, Herpesviridae Infections veterinary, Herpesvirus 1, Bovine classification
- Abstract
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 Hind III site. Our PCR-REA method is an affordable and rapid test that will subtype all BoHV-1 strains.
- Published
- 2020
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