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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.
              1. Titov I, Rutschke N, Kraft FA, Köpke M, Nebling E, Gerken M. Detection of fluorescence-labeled DNA with in-plane organic optoelectronic devices. Biomed Opt Express 2022 Dec 1;13(12):6300-6316.
                doi: 10.1364/BOE.475358pubmed: 36589587google scholar: lookup
              2. Badr C, Souiai O, Arbi M, El Behi I, Essaied MS, Khosrof I, Benkahla A, Chabchoub A, Ghram A. Epidemiological and Phylogeographic Study of Equid Herpesviruses in Tunisia. Pathogens 2022 Sep 5;11(9).
                doi: 10.3390/pathogens11091016pubmed: 36145448google scholar: lookup
              3. Carvelli A, Nielsen SS, Paillot R, Broglia A, Kohnle L. Clinical impact, diagnosis and control of Equine Herpesvirus-1 infection in Europe. EFSA J 2022 Apr;20(4):e07230.
                doi: 10.2903/j.efsa.2022.7230pubmed: 35414834google scholar: lookup
              4. Nielsen SS, Alvarez J, Bicout DJ, Calistri P, Canali E, Drewe JA, Garin-Bastuji B, Gonzales Rojas JL, Gortázar C, Herskin M, Michel V, Miranda Chueca MÁ, Roberts HC, Padalino B, Pasquali P, Spoolder H, Ståhl K, Calvo AV, Viltrop A, Winckler C, Carvelli A, Paillot R, Broglia A, Kohnle L, Baldinelli F, Van der Stede Y. Assessment of listing and categorisation of animal diseases within the framework of the Animal Health Law (Regulation (EU) No 2016/429): infection with Equine Herpesvirus-1. EFSA J 2022 Jan;20(1):e07036.
                doi: 10.2903/j.efsa.2022.7036pubmed: 35035581google scholar: lookup
              5. Samoilowa S, Giessler KS, Torres CEM, Hussey GS, Allum A, Fux R, Jerke C, Kiupel M, Matiasek K, Sledge DG, Goehring LS. Equid herpesvirus-1 Distribution in Equine Lymphoid and Neural Tissues 70 Days Post Infection. Pathogens 2021 Jun 5;10(6).
                doi: 10.3390/pathogens10060707pubmed: 34198884google scholar: lookup
              6. Zarski LM, Vaala WE, Barnett DC, Bain FT, Soboll Hussey G. A Live-Attenuated Equine Influenza Vaccine Stimulates Innate Immunity in Equine Respiratory Epithelial Cell Cultures That Could Provide Protection From Equine Herpesvirus 1. Front Vet Sci 2021;8:674850.
                doi: 10.3389/fvets.2021.674850pubmed: 34179166google scholar: lookup
              7. Zarski LM, Giessler KS, Jacob SI, Weber PSD, McCauley AG, Lee Y, Soboll Hussey G. Identification of Host Factors Associated with the Development of Equine Herpesvirus Myeloencephalopathy by Transcriptomic Analysis of Peripheral Blood Mononuclear Cells from Horses. Viruses 2021 Feb 24;13(3).
                doi: 10.3390/v13030356pubmed: 33668216google scholar: lookup
              8. Giessler KS, Samoilowa S, Soboll Hussey G, Kiupel M, Matiasek K, Sledge DG, Liesche F, Schlegel J, Fux R, Goehring LS. Viral Load and Cell Tropism During Early Latent Equid Herpesvirus 1 Infection Differ Over Time in Lymphoid and Neural Tissue Samples From Experimentally Infected Horses. Front Vet Sci 2020;7:621.
                doi: 10.3389/fvets.2020.00621pubmed: 33102556google scholar: lookup
              9. Stasiak K, Dunowska M, Rola J. Outbreak of equid herpesvirus 1 abortions at the Arabian stud in Poland. BMC Vet Res 2020 Oct 6;16(1):374.
                doi: 10.1186/s12917-020-02586-ypubmed: 33023592google scholar: lookup
              10. Pavulraj S, Kamel M, Stephanowitz H, Liu F, Plendl J, Osterrieder N, Azab W. Equine Herpesvirus Type 1 Modulates Cytokine and Chemokine Profiles of Mononuclear Cells for Efficient Dissemination to Target Organs. Viruses 2020 Sep 8;12(9).
                doi: 10.3390/v12090999pubmed: 32911663google scholar: lookup
              11. Azab W, Bedair S, Abdelgawad A, Eschke K, Farag GK, Abdel-Raheim A, Greenwood AD, Osterrieder N, Ali AAH. Detection of equid herpesviruses among different Arabian horse populations in Egypt. Vet Med Sci 2019 Aug;5(3):361-371.
                doi: 10.1002/vms3.176pubmed: 31149784google scholar: lookup
              12. Seeber PA, Dayaram A, Sicks F, Osterrieder N, Franz M, Greenwood AD. Noninvasive Detection of Equid Herpesviruses in Fecal Samples. Appl Environ Microbiol 2019 Feb 1;85(3).
                doi: 10.1128/AEM.02234-18pubmed: 30446563google scholar: lookup
              13. Ghoniem SM, El Deeb AH, Aggour MG, Hussein HA. Development and evaluation of a multiplex reverse-transcription real-time PCR assay for detection of equine respiratory disease viruses. J Vet Diagn Invest 2018 Nov;30(6):924-928.
                doi: 10.1177/1040638718799388pubmed: 30239276google scholar: lookup
              14. Costantini D, Seeber PA, Soilemetzidou SE, Azab W, Bohner J, Buuveibaatar B, Czirják GÁ, East ML, Greunz EM, Kaczensky P, Lamglait B, Melzheimer J, Uiseb K, Ortega A, Osterrieder N, Sandgreen DM, Simon M, Walzer C, Greenwood AD. Physiological costs of infection: herpesvirus replication is linked to blood oxidative stress in equids. Sci Rep 2018 Jul 9;8(1):10347.
                doi: 10.1038/s41598-018-28688-0pubmed: 29985431google scholar: lookup
              15. Dayaram A, Franz M, Schattschneider A, Damiani AM, Bischofberger S, Osterrieder N, Greenwood AD. Long term stability and infectivity of herpesviruses in water. Sci Rep 2017 Apr 21;7:46559.
                doi: 10.1038/srep46559pubmed: 28429732google scholar: lookup
              16. Houtsma A, Bedenice D, Pusterla N, Pugliese B, Mapes S, Hoffman AM, Paxson J, Rozanski E, Mukherjee J, Wigley M, Mazan MR. Association between inflammatory airway disease of horses and exposure to respiratory viruses: a case control study. Multidiscip Respir Med 2015;10:33.
                doi: 10.1186/s40248-015-0030-3pubmed: 26535117google scholar: lookup
              17. Hussey GS, Goehring LS, Lunn DP, Hussey SB, Huang T, Osterrieder N, Powell C, Hand J, Holz C, Slater J. Experimental infection with equine herpesvirus type 1 (EHV-1) induces chorioretinal lesions. Vet Res 2013 Dec 5;44(1):118.
                doi: 10.1186/1297-9716-44-118pubmed: 24308772google scholar: lookup
              18. Smith KL, Li Y, Breheny P, Cook RF, Henney PJ, Sells S, Pronost S, Lu Z, Crossley BM, Timoney PJ, Balasuriya UB. New real-time PCR assay using allelic discrimination for detection and differentiation of equine herpesvirus-1 strains with A2254 and G2254 polymorphisms. J Clin Microbiol 2012 Jun;50(6):1981-8.
                doi: 10.1128/JCM.00135-12pubmed: 22493339google scholar: lookup
              19. Soboll Hussey G, Hussey SB, Wagner B, Horohov DW, Van de Walle GR, Osterrieder N, Goehring LS, Rao S, Lunn DP. Evaluation of immune responses following infection of ponies with an EHV-1 ORF1/2 deletion mutant. Vet Res 2011 Feb 7;42(1):23.
                doi: 10.1186/1297-9716-42-23pubmed: 21314906google scholar: lookup
              20. Kambayashi Y, Bannai H, Nemoto M, Kawanishi N, Niwa H, Tsujimura K. Comparative analysis of 3 qPCR primer-probe sets for the detection of equid alphaherpesvirus 1. J Vet Diagn Invest 2026 Jan;38(1):77-83.
                doi: 10.1177/10406387251379857pubmed: 41055561google scholar: lookup
              21. Badr C, Handous M, Nsiri J, ElBehi I, Arbi M, Maaroufi A, Bennour MA, Ben Osman R, Dachraoui K, Abbes M, Mahmoudi A, Khosrof I, Abrougui S, Lachheb J, Zhioua E, Larbi I. First report of West Nile virus infections in horses in Tunisia from 2018 to 2023. Virol J 2025 Oct 1;22(1):318.
                doi: 10.1186/s12985-025-02918-0pubmed: 41035042google scholar: lookup
              22. Musoles-Cuenca B, Padilla-Blanco M, Vitale V, Lorenzo-Bermejo T, de la Cuesta-Torrado M, Ballester B, Maiques E, Rubio-Guerri C, Velloso Alvarez A. First Molecular Evidence of Equine Herpesvirus Type 1 (EHV-1) in Ocular Swabs of Clinically Affected Horses. Viruses 2025 Jun 18;17(6).
                doi: 10.3390/v17060862pubmed: 40573453google scholar: lookup
              23. Giessler KS, Goehring LS, Jacob SI, Davis A, Esser MM, Lee Y, Zarski LM, Weber PSD, Hussey GS. Impact of the host immune response on the development of equine herpesvirus myeloencephalopathy in horses. J Gen Virol 2024 May;105(5).
                doi: 10.1099/jgv.0.001987pubmed: 38767608google scholar: lookup
              24. El-Zayat M, Shemies OA, Mosad SM, El Rahman SA. Recent sequencing and phylogenetic analysis of equine herpesviruses 1 and 4 among different equine populations in Egypt. J Adv Vet Anim Res 2023 Dec;10(4):639-646.
                doi: 10.5455/javar.2023.j719pubmed: 38370903google scholar: lookup
              25. Pusterla N, Dorman DC, Burgess BA, Goehring L, Gross M, Osterrieder K, Soboll Hussey G, Lunn DP. Viremia and nasal shedding for the diagnosis of equine herpesvirus-1 infection in domesticated horses. J Vet Intern Med 2024 May-Jun;38(3):1765-1791.
                doi: 10.1111/jvim.16958pubmed: 38069548google scholar: lookup
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