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Home / Equine Research Bank / Arch Virol / Diagnosis of equine gammaherpesvirus 2 and 5 infections by p...
Archives of virology1995; 140(6); 1049-1060; doi: 10.1007/BF01315414

Diagnosis of equine gammaherpesvirus 2 and 5 infections by polymerase chain reaction.

Abstract: Nested polymerase chain reaction (PCR) assays were developed for the detection of equine herpesvirus 2 (EHV2) and equine herpesvirus 5 (EHV5) using the nucleotide sequences from the glycoprotein B (gB) gene of EHV2 and the thymidine kinase (TK) gene of EHV5. The simultaneous use of EHV2 specific and EHV5 specific primers in one nested amplification assay (multiplex PCR) enabled a rapid, specific and sensitive diagnosis for each virus. PCR was found to be 10(3) times more sensitive than virus isolation by cell culture for EHV2 and 10(6) for EHV5. In separate PCR assays, the routine detection limit after ethidium bromide staining was 0.6 fg for EHV2 plasmid DNA and 2.3 fg for EHV5 plasmid DNA, equivalent for both viruses to approximately 100 genome copies. The detection limits in multiplex PCR were 6 pg for EHV2 and 2.3 fg for EHV5, respectively. PCR assays were applied to studies of the epidemiology of EHV2 and EHV5 infections of racehorses and breeding mares in Victoria and New South Wales, Australia. Peripheral blood leukocytes from 31% of horses were positive for EHV2, 16% positive for EHV5, 8% positive for both viruses and 63% negative for both viruses. EHV2 PCR was also successfully used to detect EHV2 DNA in nasal secretions from horses. The multiplex PCR assay proved to be a rapid and reliable method for the simultaneous detection and differentiation of 2 related equine gammaherpesviruses.
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Publication Date: 1995-01-01 PubMed ID: 7611877DOI: 10.1007/BF01315414Google Scholar: Lookup
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  • Journal Article
  • Research Support
  • Non-U.S. Gov't

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.

The research article outlines the development of a more sensitive detection method for Equine Herpesvirus 2 and 5. Utilizing Nested Polymerase Chain Reaction (PCR) assays, the team improved existing diagnostic tools, allowing for earlier and more precise identification of the viruses in racehorses and breeding mares.

Nested Polymerase Chain Reaction for Detection

  • The study introduces a novel diagnostic tool developed for identifying equine herpesvirus 2 (EHV2) and equine herpesvirus 5 (EHV5), two common viruses in horses.
  • The scientists used nested polymerase chain reaction (PCR), a specific type of PCR that increases sensitivity and specificity, to identify the viruses. For this, they utilized the nucleotide sequences from specific genes of EHV2 and EHV5.
  • They modified this nested PCR into a multiplex PCR, which allows for multiple genes to be amplified at the same time. This simultaneous amplification makes the detection process quicker and more efficient.

Performance Advantage Over Previous Methods

  • Comparison tests showed that the new PCR method was far more sensitive than traditional virus isolation techniques. Specifically, it was 10^3 times more sensitive for EHV2 and 10^6 times more sensitive for EHV5.
  • In separate PCR assays, the researchers were able to detect minute amounts of the virus, effectively improving the detection limit of the disease.

Testing and Applications

  • Researchers tested their new method on racehorses and breeding mares in Victoria and New South Wales, Australia.
  • Use of the method helped them find that 31% of horses tested positive for EHV2, and 16% tested positive for EHV5. Interestingly, 8% were positive for both viruses.
  • They were also able to identify EHV2 DNA in nasal secretions from horses, which is significant as it shows the method can be used to detect the virus from various biological media, not solely blood samples.
  • The new PCR assays demonstrate significant potential for rapid and reliable detection, differentiation, and epidemiological studies involving equine gammaherpesviruses.

Cite This Article

APA
Reubel GH, Crabb BS, Studdert MJ. (1995). Diagnosis of equine gammaherpesvirus 2 and 5 infections by polymerase chain reaction. Arch Virol, 140(6), 1049-1060. https://doi.org/10.1007/BF01315414

Publication

Archives of virology
ISSN: 0304-8608
NlmUniqueID: 7506870
Country: Austria
Language: English
Volume: 140
Issue: 6
Pages: 1049-1060

Researcher Affiliations

Reubel, G H
  • Centre for Equine Virology, School of Veterinary Science, University of Melbourne, Parkville, Victoria, Australia.
Crabb, B S
    Studdert, M J

      MeSH Terms

      • Animals
      • Base Sequence
      • Cells, Cultured
      • DNA Primers
      • Gammaherpesvirinae / genetics
      • Gammaherpesvirinae / isolation & purification
      • Herpesviridae Infections / diagnosis
      • Herpesviridae Infections / epidemiology
      • Herpesviridae Infections / veterinary
      • Herpesviridae Infections / virology
      • Horse Diseases / diagnosis
      • Horse Diseases / epidemiology
      • Horse Diseases / virology
      • Horses
      • Leukocytes, Mononuclear / virology
      • Molecular Sequence Data
      • Nasal Mucosa / virology
      • Polymerase Chain Reaction / methods
      • Polymerase Chain Reaction / veterinary
      • Sensitivity and Specificity

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      Citations

      This article has been cited 7 times.
      1. 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).
        doi: 10.3390/ani11123418pubmed: 34944194google scholar: lookup
      2. Thorsteinsdóttir L, Jónsdóttir S, Stefánsdóttir SB, Andrésdóttir V, Wagner B, Marti E, Torsteinsdóttir S, Svansson V. The effect of maternal immunity on the equine gammaherpesvirus type 2 and 5 viral load and antibody response.. PLoS One 2019;14(6):e0218576.
        doi: 10.1371/journal.pone.0218576pubmed: 31226153google scholar: lookup
      3. Dall Agnol AM, Beuttemmuller EA, Pilz D, Leme RA, Saporiti V, Headley SA, Alfieri AF, Alfieri AA. Detection of Equid gammaherpesvirus 2 and 5 DNA in the upper respiratory tract of asymptomatic horses from Southern Brazil.. Braz J Microbiol 2019 Jul;50(3):875-878.
        doi: 10.1007/s42770-019-00100-7pubmed: 31187445google scholar: lookup
      4. Marenzoni ML, Stefanetti V, Danzetta ML, Timoney PJ. Gammaherpesvirus infections in equids: a review.. Vet Med (Auckl) 2015;6:91-101.
        doi: 10.2147/VMRR.S39473pubmed: 30155436google scholar: lookup
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        doi: 10.1128/CMR.13.4.559pubmed: 11023957google scholar: lookup
      7. Drummer HE, Reubel GH, Studdert MJ. Equine gammaherpesvirus 2 (EHV2) is latent in B lymphocytes.. Arch Virol 1996;141(3-4):495-504.
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