Equine herpesvirus-4 kinetics in peripheral blood leukocytes and nasopharyngeal secretions in foals using quantitative real-time TaqMan PCR.
Abstract: Based on the hypothesis that the viral load of cells infected with EHV-4 will likely change during the course of disease, TaqMan PCR was used to investigate and characterize the kinetics of EHV-4 viral DNA load (glycoprotein B gene) and transcriptional activity (glycoprotein B and latency-associated transcripts) in peripheral blood leukocytes (PBLs) and nasopharyngeal secretions (NSs) collected from 11 foals during a field outbreak of respiratory disease. The EHV-4 DNA load in PBLs was low and of short duration after onset of clinical signs. In contrast, the EHV-4 load in NSs remained high for the majority of the foals over a period of 4 weeks. Viral replication determined by detection of mRNA expression of the structural glycoprotein B was detected only in NSs during the first 7 days after onset of clinical signs for most foals. The majority of foals expressed latency-associated transcripts in NS sonly during the first 7 days after onset of clinical signs. Persistence of the expression of latency-associated transcripts in NS, as a reflection of a latent viral state, was not documented during the 28-day study period. Based on these results, it was concluded that lytic infection with EHV-4 can be diagnosed either by high EHV-4 DNA load of glycoprotein B gene or by detection of transcriptional activity of glycoprotein B.
Publication Date: 2006-02-16 PubMed ID: 16475518DOI: 10.1177/104063870501700610Google Scholar: Lookup
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- Journal Article
- Research Support
- Non-U.S. Gov't
Summary
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The research article investigates the progression of Equine herpesvirus-4 (EHV-4) infection in foals through the analysis of viral DNA load and transcriptional activity in blood leukocytes and nasopharyngeal secretions, using a technique known as TaqMan PCR.
Overview of Study Design:
- The study was designed to explore the hypothesis that the viral load of cells infected with EHV-4 changes over the course of the disease.
- The researchers used TaqMan real-time PCR to examine and characterize EHV-4 viral DNA load (specifically the glycoprotein B gene) and transcriptional activity (the glycoprotein B and latency-associated transcripts).
- Data for the study was collected from peripheral blood leukocytes (PBLs) and nasopharyngeal secretions (NSs) collected from 11 foals during an outbreak of respiratory disease.
Findings of the Study:
- Findings showed that EHV-4 DNA load in PBLs was low and lasted a short duration following the onset of clinical signs. Unlike this, the EHV-4 load in NSs remained high over a period of 4 weeks for the majority of the foals.
- Evidence of viral replication (based on the detection of mRNA expression of the structural glycoprotein B) was only found in NSs during the first week of disease onset for most foals.
- Most of the foals only expressed latency-associated transcripts (an indication of dormant virus) in their NSs during the first week after disease onset. The study observed no persistence of this expression, showing no long-term dormant viral state, through the 28-day study period.
Conclusions of the Study:
- The study concludes that lytic infection (active viral replication leading to cell death) with EHV-4 can be diagnosed either by high EHV-4 DNA load of the glycoprotein B gene, or by detection of transcriptional activity of the glycoprotein B.
- These findings contribute to our understanding of EHV-4 infection kinetics and can guide future research and clinical efforts towards effectively diagnosing and managing the disease.
Cite This Article
APA
Pusterla N, Leutenegger CM, Wilson WD, Watson JL, Ferraro GL, Madigan JE.
(2006).
Equine herpesvirus-4 kinetics in peripheral blood leukocytes and nasopharyngeal secretions in foals using quantitative real-time TaqMan PCR.
J Vet Diagn Invest, 17(6), 578-581.
https://doi.org/10.1177/104063870501700610 Publication
Researcher Affiliations
- Department of Medicine and Epidemiology, School of Veterinary Medicine, University of California, Davis 95616, USA.
MeSH Terms
- Animals
- California / epidemiology
- Disease Outbreaks / veterinary
- Female
- Herpesviridae Infections / epidemiology
- Herpesviridae Infections / veterinary
- Herpesviridae Infections / virology
- Herpesvirus 4, Equid / genetics
- Herpesvirus 4, Equid / isolation & purification
- Horse Diseases / diagnosis
- Horse Diseases / epidemiology
- Horse Diseases / virology
- Horses / blood
- Horses / virology
- Leukocytes / virology
- Male
- Mucus / metabolism
- Mucus / virology
- Nasopharynx / metabolism
- Nasopharynx / virology
- Polymerase Chain Reaction / veterinary
- Virus Replication
Citations
This article has been cited 14 times.- Mureşan A, Mureşan C, Siteavu M, Avram E, Bochynska D, Taulescu M. An Outbreak of Equine Herpesvirus-4 in an Ecological Donkey Milk Farm in Romania. Vaccines (Basel) 2022 Mar 18;10(3).
- El-Hage C, Mekuria Z, Dynon K, Hartley C, McBride K, Gilkerson J. Association of Equine Herpesvirus 5 with Mild Respiratory Disease in a Survey of EHV1, -2, -4 and -5 in 407 Australian Horses. Animals (Basel) 2021 Nov 30;11(12).
- Pavulraj S, Eschke K, Theisen J, Westhoff S, Reimers G, Andreotti S, Osterrieder N, Azab W. Equine Herpesvirus Type 4 (EHV-4) Outbreak in Germany: Virological, Serological, and Molecular Investigations. Pathogens 2021 Jun 25;10(7).
- Salco R, Bowers J, Hernandez V, Barnum S, Pusterla N. Prevention of respiratory infections with alpha- and gamma-herpesviruses in weanling foals by using a modified live intra-nasal equine influenza vaccine. Can Vet J 2020 May;61(5):517-520.
- 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.
- Holz CL, Sledge DG, Kiupel M, Nelli RK, Goehring LS, Soboll Hussey G. Histopathologic Findings Following Experimental Equine Herpesvirus 1 Infection of Horses. Front Vet Sci 2019;6:59.
- Taktaz Hafshejani T, Nekoei S, Vazirian B, Doosti A, Khamesipour F, Anyanwu MU. Molecular Detection of Equine Herpesvirus Types 1 and 4 Infection in Healthy Horses in Isfahan Central and Shahrekord Southwest Regions, Iran. Biomed Res Int 2015;2015:917854.
- Spiesschaert B, Goldenbogen B, Taferner S, Schade M, Mahmoud M, Klipp E, Osterrieder N, Azab W. Role of gB and pUS3 in Equine Herpesvirus 1 Transfer between Peripheral Blood Mononuclear Cells and Endothelial Cells: a Dynamic In Vitro Model. J Virol 2015 Dec;89(23):11899-908.
- Azab W, Osterrieder N. Glycoproteins D of equine herpesvirus type 1 (EHV-1) and EHV-4 determine cellular tropism independently of integrins. J Virol 2012 Feb;86(4):2031-44.
- Ji Y, Xu D, Si W, Zhang Y, Khan MZ, Zhao X, Liu W. Transcriptomic and Proteomic Profiling of Rabbit Kidney Cells Infected with Equine Herpesvirus 8. Viruses 2025 Apr 29;17(5).
- Pusterla N, Barnum S, Lawton K, Craig B, James K. Investigation of the frequency and selected prevalence factors of equid alphaherpesvirus 4 viremia in horses with acute onset of fever and respiratory signs. J Vet Diagn Invest 2025 Mar;37(2):349-353.
- Charbonnel A, Lavoie JP, Juette T, St-Sauveur VG, Denis S, Gagnon CA, Leclère M. Comparison of nasal swabs and handmade foam cubes for detecting equine herpesvirus 5 (EHV-5) by quantitative polymerase chain reaction (qPCR). Can J Vet Res 2025 Jan;89(1):11-16.
- Ryt-Hansen P, Johansen VK, Cuicani MM, Larsen LE, Hansen S. Outbreak of equine herpesvirus 4 (EHV-4) in Denmark: tracing patient zero and viral characterization. BMC Vet Res 2024 Jul 3;20(1):287.
- Lebrasseur O, More KD, Orlando L. Equine herpesvirus 4 infected domestic horses associated with Sintashta spoke-wheeled chariots around 4,000 years ago. Virus Evol 2024;10(1):vead087.
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