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A novel PCR protocol for detection and differentiation of neuropathogenic and non-neuropathogenic equid alphaherpesvirus 1.
- Source :
-
Journal of veterinary diagnostic investigation : official publication of the American Association of Veterinary Laboratory Diagnosticians, Inc [J Vet Diagn Invest] 2019 Sep; Vol. 31 (5), pp. 696-703. - Publication Year :
- 2019
-
Abstract
- Equid alphaherpesvirus 1 (EHV-1) infections can have a major impact on the horse industry and equine welfare by causing abortion or respiratory or neurologic disease. A single nucleotide polymorphism (A <subscript>2254</subscript> →G <subscript>2254</subscript> ) in open reading frame (ORF) 30, encoding the catalytic subunit of the DNA polymerase, has been shown to be a strong predictive marker for neuropathogenicity. Given that a previously established real-time PCR (rtPCR) protocol yielded unsatisfactory results concerning determination of the EHV-1 genotype, we developed and evaluated a new conventional PCR protocol enabling identification of the genotype by sequencing and restriction enzyme analysis (REA). Thirty samples from horses with signs typical for EHV-1 infection were tested by rtPCR and our new conventional PCR. The results showed that compared to rtPCR, the conventional PCR protocol combined with sequencing and REA was more reliable concerning unambiguous determination of the EHV-1 genotype. Results of our new assay confirmed previous findings, according to which the non-neuropathogenic genotype A <subscript>2254</subscript> is predominantly found in animals with fever, respiratory signs, and abortions or perinatal mortality, whereas the neuropathogenic genotype G <subscript>2254</subscript> is primarily detected in animals suffering from neurologic disease. In some samples, results pointed towards coinfection with both genotypes. Further studies are required in order to elucidate the significance of infections with genotype A <subscript>2254</subscript> and G <subscript>2254</subscript> in neurologic and non-neurologic cases, respectively.
- Subjects :
- Animals
DNA-Directed DNA Polymerase genetics
Female
Genotype
Herpesviridae Infections virology
Herpesvirus 1, Equid genetics
Horses
Nervous System Diseases virology
Open Reading Frames
Pregnancy
Real-Time Polymerase Chain Reaction methods
Restriction Mapping veterinary
Herpesviridae Infections veterinary
Herpesvirus 1, Equid classification
Horse Diseases virology
Nervous System Diseases veterinary
Polymorphism, Single Nucleotide
Real-Time Polymerase Chain Reaction veterinary
Subjects
Details
- Language :
- English
- ISSN :
- 1943-4936
- Volume :
- 31
- Issue :
- 5
- Database :
- MEDLINE
- Journal :
- Journal of veterinary diagnostic investigation : official publication of the American Association of Veterinary Laboratory Diagnosticians, Inc
- Publication Type :
- Academic Journal
- Accession number :
- 31477001
- Full Text :
- https://doi.org/10.1177/1040638719871975