Characterization of viral loads, strain and state of equine herpesvirus-1 using real-time PCR in horses following natural exposure at a racetrack in California

Abstract

The objective of this study was to determine viral loads, strain (neuropathogenic versus non-neuropathogenic) and state (lytic, non-replicating, latent) of equine herpesvirus-1 (EHV-1) by real-time polymerase chain reaction (PCR) in the blood and nasopharyngeal secretions of adult horses following natural exposure. The index case, a 4-year-old Thoroughbred gelding with confirmed EHV-1 myeloencephalopathy, as well as potentially exposed horses, were sampled over a period of 3 weeks. The study population comprised of 39 adult Thoroughbred horses and 35 adult “pony” and outrider horses of various breeds housed at a racetrack in Northern California. Blood samples and nasopharyngeal secretions (NPS) from all horses were tested on several occasions for EHV-1 DNA viral loads, targeting the glycoprotein B ( gB) gene, viral strain, targeting the ORF 30 gene, and transcriptional activity of EHV-1, targeting the gB gene and latency-associated transcripts (LATs). Viral loads and transcriptional activity of the gB gene declined rapidly in the index case following antiviral treatment. The prevalence of EHV-1 infection in NPS determined by PCR slowly decreased over the 22 day study period from 25% to 14%. The initial surveillance showed multiple clusters of exposure, one associated with the index case and two related to horses that had recently returned from a different racetrack. Viral strain differentiation showed that only two horses (the index case and a neighboring horse) were infected with only a neuropathogenic strain, while all other horses were infected with either a non-neuropathogenic strain or were dually infected with both neuropathogenic and non-neuropathogenic strains. In most cases, the virus was present in either a lytic or a non-replicating form, while latent virus was found in blood and NPS much less frequently. The molecular approach used in this study showed promise for assessing the risk of exposing other horses to EHV-1 and for studying viral kinetics in infected horses.