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Home / Equine Research Bank / J Vet Diagn Invest / Detection and quantification of equine herpesvirus-1 viremia...
Journal of veterinary diagnostic investigation : official publication of the American Association of Veterinary Laboratory Diagnosticians, Inc2006; 18(4); 335-342; doi: 10.1177/104063870601800403

Detection and quantification of equine herpesvirus-1 viremia and nasal shedding by real-time polymerase chain reaction.

Abstract: Equine herpesvirus-1 (EHV-1) infection is common in young horses throughout the world, resulting in respiratory disease, epidemic abortion, sporadic myelitis, or latent infections. To improve on conventional diagnostic tests for EHV-1, a real-time polymerase chain reaction (PCR) technique was developed, using primers and probes specific for the EHV-1 gB gene. Amplification efficiencies of 100% +/- 5% were obtained for DNA isolated from a plasmid, infected peripheral blood mononuclear cells (PBMCs), and nasal secretions from infected ponies. The dynamic range of the assay was 8 log10 dilutions, and the lower limit of detection was 6 DNA copies. Fifteen ponies, seronegative for EHV-1, were experimentally infected with EHV-1, and nasal samples were used to quantify shedding of virus by both virus isolation and real-time PCR analysis. Virus isolation identified nasal shedding of EHV-1 in 12/15 ponies on a total of 25 days; real-time PCR detected viral shedding in 15/15 ponies on 75 days. Viremia was quantified using PBMC DNA, subsequent to challenge infection in 3 additional ponies. Viremia was identified in 1/3 ponies on a single day by virus isolation; real-time PCR detected viremia in 3/3 ponies on 17 days. When real-time PCR was used to analyze PBMC DNA from 11 latently infected ponies (documented by nested PCR), EHV-1 was not detected. We conclude that real-time PCR is a sensitive and quantitative test for EHV-1 nasal shedding and viremia and provides a valuable tool for EHV-1 surveillance, diagnosis of clinical disease, and investigation of vaccine efficacy.
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Publication Date: 2006-08-23 PubMed ID: 16921871DOI: 10.1177/104063870601800403Google Scholar: Lookup
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  • Journal Article
  • Research Support
  • Non-U.S. Gov't
  • Research Support
  • U.S. Gov't
  • Non-P.H.S.

Summary

This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.

This research work attempted to develop a sensitive and reliable method for detecting and quantifying Equine Herpesvirus-1 (EHV-1) in horses using a real-time polymerase chain reaction (PCR) technique. Results of the study showed that real-time PCR was able to detect EHV-1 not only in nasal samples but also in the blood of infected animals, making it a potent tool in managing and controlling EHV-1 occurrence.

Introduction and Methodology

  • The EHV-1 infection occurs worldwide in young horses, resulting in different conditions such as respiratory diseases, abortions, sporadic myelitis, and latent infections.
  • The study sought to improve the standard diagnostic tests for EHV-1. This was achieved by developing a real-time PCR technique that utilizes primers and probes specific to the EHV-1 gB gene.
  • The efficacy of the amplification process was tested with DNA sourced from infected plasmids, peripheral blood mononuclear cells (PBMCs), and the nasal secretions from infected ponies. The results showed a 100% efficiency with a 5% margin for error.
  • 15 ponies that were not seropositive for EHV-1 were deliberately exposed to the virus. Nasal samples from these ponies were then used to identify and quantify the virus’s shedding.

Results

  • The virus isolation process was able to identify EHV-1 shedding in 12 out of the 15 ponies across a total of 25 days.
  • Real-time PCR, on the other hand, was able to detect viral shedding in all 15 ponies over a period of 75 days dictating a higher sensitivity of this method.
  • Blood samples from 3 additional ponies were tested using the same methods after exposing them to EHV-1. Virus isolation identified viremia in one pony for a single day, while real-time PCR was able to identify viremia in all three ponies across 17 days.
  • When real-time PCR was used to analyze PBMC DNA from 11 ponies thought to be latently carrying EHV-1, the virus wasn’t detected at all.

Conclusion

  • The findings of this study suggest that real-time PCR is a highly sensitive and quantitative test for EHV-1 nasal shedding and viremia.
  • This makes it a valuable tool for EHV-1 disease surveillance, clinical diagnosis, and the evaluation of vaccine effectiveness against the virus.

Cite This Article

APA
Hussey SB, Clark R, Lunn KF, Breathnach C, Soboll G, Whalley JM, Lunn DP. (2006). Detection and quantification of equine herpesvirus-1 viremia and nasal shedding by real-time polymerase chain reaction. J Vet Diagn Invest, 18(4), 335-342. https://doi.org/10.1177/104063870601800403

Publication

Journal of veterinary diagnostic investigation : official publication of the American Association of Veterinary Laboratory Diagnosticians, Inc
ISSN: 1040-6387
NlmUniqueID: 9011490
Country: United States
Language: English
Volume: 18
Issue: 4
Pages: 335-342

Researcher Affiliations

Hussey, Stephen B
  • Department of Clinical Sciences, College of Veterinary Medicine and Biomedical Sciences, Colorado State University, 300 West Drake Road, Fort Collins, CO 80523, USA.
Clark, Rodney
    Lunn, Katharine F
      Breathnach, Cormac
        Soboll, Gisela
          Whalley, J Millar
            Lunn, D Paul

              MeSH Terms

              • Animals
              • Female
              • Herpesvirus 1, Equid / genetics
              • Herpesvirus 1, Equid / isolation & purification
              • Horse Diseases / diagnosis
              • Horse Diseases / virology
              • Horses / virology
              • Male
              • Polymerase Chain Reaction / methods
              • Polymerase Chain Reaction / veterinary
              • Sensitivity and Specificity
              • Viremia / veterinary
              • Viremia / virology
              • Virus Latency
              • Virus Shedding

              Citations

              This article has been cited 25 times.
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