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Journal of virological methods2005; 131(1); 92-98; doi: 10.1016/j.jviromet.2005.07.010

Detection of equine herpesvirus type 1 using a real-time polymerase chain reaction.

Abstract: Equid herpesvirus 1 (EHV1) is a major disease of equids worldwide causing considerable losses to the horse industry. A variety of techniques, including PCR have been used to diagnose EHV1. Some of these PCRs were used in combination with other techniques such as restriction enzyme analysis (REA) or hybridisation, making them cumbersome for routine diagnostic testing and increasing the chances of cross-contamination. Furthermore, they involve the use of suspected carcinogens such as ethidium bromide and ultraviolet light. In this paper, we describe a real-time PCR, which uses minor groove-binding probe (MGB) technology for the diagnosis of EHV1. This technique does not require post-PCR manipulations thereby reducing the risk of cross-contamination. Most importantly, the technique is specific; it was able to differentiate EHV1 from the closely related member of the Alphaherpesvirinae, equid herpesvirus 4 (EHV4). It was not reactive with common opportunistic pathogens such as Escherichia coli, Klebsiella oxytoca, Pseudomonas aeruginosa and Enterobacter agglomerans often involved in abortion. Similarly, it did not react with equine pathogens such as Streptococcus equi, Streptococcus equisimilis, Streptococcus zooepidemicus, Taylorella equigenitalis and Rhodococcus equi, which also cause abortion. The results obtained with this technique agreed with results from published PCR methods. The assay was sensitive enough to detect EHV1 sequences in paraffin-embedded tissues and clinical samples. When compared to virus isolation, the test was more sensitive. This test will be useful for the routine diagnosis of EHV1 based on its specificity, sensitivity, ease of performance and rapidity.
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Publication Date: 2005-08-30 PubMed ID: 16137772DOI: 10.1016/j.jviromet.2005.07.010Google Scholar: Lookup
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  • Journal Article

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 deals with the detection method inside of an improved Polymerase Chain Reaction (PCR) test for Equid Herpesvirus 1 (EHV1), a significantly damaging disease to horses across the globe.

Background

  • The focus of this research is Equid herpesvirus 1 (EHV1), a major disease affecting horses worldwide that results in significant economic damage in the equine industry. EHV1 has been diagnosed using a variety of techniques, including the Polymerase Chain Reaction (PCR), a standard laboratory method that is usually used in tandem with other methods such as restriction enzyme analysis (REA) or hybridisation.
  • However, these combined techniques are complex for routine diagnostic testing and increase the possibility of cross-contamination. Additionally, they involve using potentially carcinogenic substances like ethidium bromide and ultraviolet light. This paper introduces a real-time PCR with a minor groove-binding probe that offers a safer, effective alternative.

Proposed Technique

  • This new PCR approach mentioned in the paper eliminates the need for post-PCR manipulations, reducing the risk of cross-contamination. It also makes use of minor groove-binding probe (MGB) technology that guarantees a more accurate diagnosis of EHV1.
  • One of the key benefits of this technique is its specificity. It was successful in differentiating EHV1 from other closely related viruses like EHV4. It also did not give false positives with common pathogens such as E. coli, Klebsiella oxytoca, Pseudomonas aeruginosa and Enterobacter agglomerans that are usually involved in horse abortions.
  • Similarly, the technique did not react with equine pathogens including Streptococcus equi, Streptococcus equisimilis, Streptococcus zooepidemicus, Taylorella equigenitalis and Rhodococcus equi, all of which could also result in abortions for horses. Thus, it proved its value in maintaining a high degree of specificity and ensuring accurate diagnosis.

Validation and Comparison

  • The results acquired from this technique were found to be in agreement with results from already published PCR methods, making it a reliable method for testing.
  • The sensitivity of this new PCR test was high enough to detect EHV1 sequences even in paraffin-embedded tissues and other clinical samples. This high sensitivity makes the test a valuable tool for accurate and early detection of EHV1.
  • When compared with traditional virus isolation methods, this improved PCR process was found to be more sensitive.

Conclusion

  • The new test method mentioned in the paper offers a substantial improvement in terms of specificity, sensitivity, ease of use, and speed. These advantages make it a promising tool for routine diagnosis of EHV1 in horses, helping to manage and control this damaging equine disease more effectively.

Cite This Article

APA
Diallo IS, Hewitson G, Wright L, Rodwell BJ, Corney BG. (2005). Detection of equine herpesvirus type 1 using a real-time polymerase chain reaction. J Virol Methods, 131(1), 92-98. https://doi.org/10.1016/j.jviromet.2005.07.010

Publication

Journal of virological methods
ISSN: 0166-0934
NlmUniqueID: 8005839
Country: Netherlands
Language: English
Volume: 131
Issue: 1
Pages: 92-98

Researcher Affiliations

Diallo, Ibrahim S
  • Animal Research Institute, Yeerongpilly Veterinary Laboratory, Department of Primary Industries and Fisheries, Locked Mail Bag 5, Moorooka, Qld 4105, Australia. ibrahim.diallo@dpi.qld.gov.au
Hewitson, Glen
    Wright, Lucia
      Rodwell, Barry J
        Corney, Bruce G

          MeSH Terms

          • Animals
          • Genes, Viral / genetics
          • Herpesviridae Infections / diagnosis
          • Herpesviridae Infections / veterinary
          • Herpesvirus 1, Equid / genetics
          • Herpesvirus 1, Equid / isolation & purification
          • Horse Diseases / diagnosis
          • Horses
          • Oxadiazoles
          • Polymerase Chain Reaction / methods
          • Sensitivity and Specificity
          • Species Specificity
          • Viral Envelope Proteins / genetics
          • Viral Proteins

          Citations

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