Lechmann J Schoster A Ernstberger M Fouché N Fraefel C Bachofen C 2019 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. application/pdf https://sonar.ch/global/documents/235663 eng info:eu-repo/semantics/altIdentifier/doi/10.1177/1040638719871975 info:eu-repo/semantics/altIdentifier/pmid/31477001 info:eu-repo/semantics/openAccess Journal of veterinary diagnostic investigation : official publication of the American Association of Veterinary Laboratory Diagnosticians, Inc. - 2019 PCR equid alphaherpesvirus 1 neuropathogenic restriction enzyme analysis sequencing Animals DNA-Directed DNA Polymerase Female Genotype Herpesviridae Infections Herpesvirus 1, Equid Horse Diseases Horses Nervous System Diseases Open Reading Frames Polymorphism, Single Nucleotide Pregnancy Real-Time Polymerase Chain Reaction Restriction Mapping A novel PCR protocol for detection and differentiation of neuropathogenic and non-neuropathogenic equid alphaherpesvirus 1. http://purl.org/coar/resource_type/c_6501