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A novel PCR protocol for detection and differentiation of neuropathogenic and non-neuropathogenic equid alphaherpesvirus 1.

Authors
  • Lechmann, Julia1, 2
  • Schoster, Angelika1, 2
  • Ernstberger, Martina1, 2
  • Fouché, Nathalie1, 2
  • Fraefel, Cornel1, 2
  • Bachofen, Claudia1, 2
  • 1 Institute of Virology (Lechmann, Fraefel, Bachofen), Equine Department, Clinic for Equine Internal Medicine (Schoster), Department of Farm Animals, Division of Herd Medicine and Outpatient Clinic (Ernstberger), Vetsuisse Faculty, University of Zurich, Zurich Switzerland. , (Switzerland)
  • 2 Swiss Institute of Equine Medicine ISME, Department of Clinical Veterinary Medicine, Vetsuisse Faculty, University of Bern, and Agroscope, Bern, Switzerland (Fouché). , (Switzerland)
Type
Published Article
Journal
Journal of veterinary diagnostic investigation : official publication of the American Association of Veterinary Laboratory Diagnosticians, Inc
Publication Date
Sep 01, 2019
Volume
31
Issue
5
Pages
696–703
Identifiers
DOI: 10.1177/1040638719871975
PMID: 31477001
Source
Medline
Keywords
Language
English
License
Unknown

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 (A2254→G2254) 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 A2254 is predominantly found in animals with fever, respiratory signs, and abortions or perinatal mortality, whereas the neuropathogenic genotype G2254 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 A2254 and G2254 in neurologic and non-neurologic cases, respectively.

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