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Veterinary microbiology2006; 121(1-2); 18-28; doi: 10.1016/j.vetmic.2006.11.009

Detection of respiratory herpesviruses in foals and adult horses determined by nested multiplex PCR.

Abstract: A nested multiplex PCR was developed as a rapid (<12h), sensitive test for the simultaneous identification of equine herpesviruses (EHV1, EHV4, EHV2 and EHV5) in clinical samples from horses. Peripheral blood and nasal swab (NS) samples from 205 weanling Thoroughbred foals on 6 different studs over 3 consecutive seasons and from 92 adult horses without clinical signs of respiratory disease were examined using direct multiplex PCR of clinical samples (direct PCR) and conventional cell culture with differentiation of EHV in cell cultures by multiplex PCR. Multiplex PCR proved a sensitive and specific technique for the detection of EHV in cell culture and clinical samples. The technique described appeared equally sensitive as one using a single set of primers for individual EHV but reduced labour and reagent costs. Cell cultures showing cytopathic effect (CPE) were always positive for EHV on PCR. EHV were also detected by multiplex PCR in 11 samples which failed to show CPE. By a combination of multiplex PCR and cell culture or direct multiplex PCR, the presence of up to three EHV in the same sample was detected. Overall, EHV5 was detected by direct multiplex PCR of peripheral blood mononuclear cells (PBMC) and/or NS samples from 78% of foals and 47% of adult horses. Repeated sampling or cell culture in combination with multiplex PCR and with the incorporation of IL-2 in culture medium increased the sensitivity for detection of EHV in PBMC and demonstrated that EHV5 DNA could be identified in PBMC from 89% of foals and 100% of adult horses. EHV2 was identified from approximately 30% of foals, but was more frequently identified in samples from 17 foals with mild respiratory disease and was isolated infrequently from adult horses. EHV1 and EHV4 were identified uncommonly in any population in the current study.
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Publication Date: 2006-11-21 PubMed ID: 17208393DOI: 10.1016/j.vetmic.2006.11.009Google Scholar: Lookup
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  • Comparative Study
  • 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 study employed a fast and sensitive testing method known as nested multiplex PCR for the simultaneous identification of four types of equine herpesviruses (EHV) in clinical samples from horses. This technique proved more cost-effective and labor-efficient compared to utilizing a single set of primers for distinct EHVs. The prevalence of different EHVs was also determined in weanling Thoroughbred foals and adult horses, revealing varying rates of detection among different viruses and age groups.

Methodology

  • The nested multiplex PCR was a test method used for identifying EHV1, EHV4, EHV2, and EHV5 in horse samples. It was noted for its speedy results and high sensitivity levels.
  • Clinical samples, involving peripheral blood and nasal swabs, were collected from 205 weanling Thoroughbred foals across six studs for three seasons as well as from 92 adult horses showing no clinical signs of respiratory disease.
  • The researchers used both the direct multiplex PCR of clinical samples and conventional cell culture followed by differentiation of EHV in cell cultures using multiplex PCR.

Results

  • The use of the multiplex PCR method yielded sensitive and specific results in detecting EHV from both cell cultures and clinical samples.
  • Samples that revealed a cytopathic effect were consistently found to be positive for EHV through PCR testing methods.
  • Multiplex PCR and combinations with cell culture and direct multiplex PCR unveiled the presence of up to three different EHVs in the same sample.
  • Overall, EHV5 was detected in 78% of foal peripheral blood mononuclear cells or nasal swabs and 47% of adult horse samples via the direct multiplex PCR method.
  • Repeated sampling or cell culture along with the incorporation of IL-2 in culture mediums increased the sensitivity of EHV detection in peripheral blood mononuclear cells and demonstrated the identification of EHV5 DNA in 89% of foals and 100% of adult horses.
  • EHV2 was identified in about 30% of foal samples but was found more frequently in samples from 17 foals with mild respiratory disease. However, it was rarely identified in adult horse samples.
  • EHV1 and EHV4 were rarely detected in any of the studied population.

Cite This Article

APA
Wang L, Raidal SL, Pizzirani A, Wilcox GE. (2006). Detection of respiratory herpesviruses in foals and adult horses determined by nested multiplex PCR. Vet Microbiol, 121(1-2), 18-28. https://doi.org/10.1016/j.vetmic.2006.11.009

Publication

Veterinary microbiology
ISSN: 0378-1135
NlmUniqueID: 7705469
Country: Netherlands
Language: English
Volume: 121
Issue: 1-2
Pages: 18-28

Researcher Affiliations

Wang, L
  • Division of Veterinary and Biomedical Sciences, Murdoch University, Murdoch, WA 6150, Australia.
Raidal, S L
    Pizzirani, A
      Wilcox, G E

        MeSH Terms

        • Animals
        • DNA, Viral / chemistry
        • DNA, Viral / genetics
        • Herpesviridae / genetics
        • Herpesviridae / isolation & purification
        • Herpesviridae Infections / blood
        • Herpesviridae Infections / veterinary
        • Herpesviridae Infections / virology
        • Horse Diseases / diagnosis
        • Horse Diseases / virology
        • Horses
        • Polymerase Chain Reaction / methods
        • Polymerase Chain Reaction / veterinary
        • Sensitivity and Specificity

        Citations

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