Detection of equine herpesvirus type 1 by real time PCR.
Abstract: A real-time PCR assay was developed for detection and quantitation of equid herpesvirus type 1 (EHV-1). The sensitivity of the assay was compared with an established nested-PCR (n-PCR). The real-time PCR detected 1 copy of target DNA, with a sensitivity 1 log higher than gel-based n-PCR. The assay was able to detect specifically EHV-1 DNA in equine tissue samples and there was no cross-amplification of other horse herpesviruses. Real-time PCR was applied to determine EHV-1 load in tissue samples from equine aborted fetuses. The high sensitivity and reproducibility of the EHV-1-specific fluorogenic PCR assay, combined with the wide dynamic range and the high throughput, make this method suitable for diagnostic and research applications.
Publication Date: 2005-11-22 PubMed ID: 16309751DOI: 10.1016/j.jviromet.2005.10.024Google Scholar: Lookup
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- Journal Article
Summary
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This research discusses the development of a real-time PCR assay for the detection and quantification of equid herpesvirus type 1, an equine virus. The study reveals that this method is notably more sensitive, wide-ranging and reproducible than previous detection methods, such as nested-PCR, making it highly suitable for both diagnostic and research applications.
Development and Testing of the Real-Time PCR Assay
- The researchers in the study developed a real-time PCR assay for the precise detection and quantification of equid herpesvirus type 1 (EHV-1).
- This assay was tested for its sensitivity and compared with an existing detection method called nested-PCR (n-PCR). The comparison showed that the real-time PCR assay was significantly more sensitive, able to detect even a single copy of target DNA, making it 1 log more sensitive than the gel-based n-PCR.
Specificity of the Real-Time PCR Assay for EHV-1
- Importantly, the assay was found to be specifically able to detect EHV-1 DNA in equine tissue samples.
- The research demonstrated that this assay did not give false positives for other types of horse herpesviruses, confirming its specificity for detecting EHV-1. This means that the assay does not cross-amplify other horse herpesviruses, thus reducing the chance of false-positive results.
Applicability of the Real-Time PCR Assay
- The real-time PCR assay was also applied successfully to measure the load of EHV-1 in tissue samples from equine aborted fetuses.
- Such sensitivity and reproducibility of this EHV-1-specific real-time PCR assay, combined with its wide dynamic range and capacity for high throughput, make it an ideal method for many diagnostic applications, where fast and accurate detection of EHV-1 is crucial.
- In addition to its diagnostic applications, the real-time PCR assay can also be effectively used in research settings where detailed, quantitive measurement of EHV-1 is required.
Cite This Article
APA
Elia G, Decaro N, Martella V, Campolo M, Desario C, Lorusso E, Cirone F, Buonavoglia C.
(2005).
Detection of equine herpesvirus type 1 by real time PCR.
J Virol Methods, 133(1), 70-75.
https://doi.org/10.1016/j.jviromet.2005.10.024 Publication
Researcher Affiliations
- Department of Animal Health and Well-being, Faculty of Veterinary Medicine of Bari, 70010 Valenzano, Bari, Italy. g.elia@veterinaria.uniba.it
MeSH Terms
- Aborted Fetus / virology
- Animals
- DNA, Viral / analysis
- DNA, Viral / genetics
- Female
- Gene Dosage
- Herpesviridae Infections / veterinary
- Herpesviridae Infections / virology
- Herpesvirus 1, Equid / genetics
- Horse Diseases / virology
- Horses
- Pregnancy
- Reproducibility of Results
- Reverse Transcriptase Polymerase Chain Reaction / veterinary
- Sensitivity and Specificity
- Viral Load
Citations
This article has been cited 11 times.- 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.
- Schnabel CL, Babasyan S, Rollins A, Freer H, Wimer CL, Perkins GA, Raza F, Osterrieder N, Wagner B. An Equine Herpesvirus Type 1 (EHV-1) Ab4 Open Reading Frame 2 Deletion Mutant Provides Immunity and Protection from EHV-1 Infection and Disease. J Virol 2019 Nov 15;93(22).
- Wimer CL, Schnabel CL, Perkins G, Babasyan S, Freer H, Stout AE, Rollins A, Osterrieder N, Goodman LB, Glaser A, Wagner B. The deletion of the ORF1 and ORF71 genes reduces virulence of the neuropathogenic EHV-1 strain Ab4 without compromising host immunity in horses. PLoS One 2018;13(11):e0206679.
- Schnabel CL, Wimer CL, Perkins G, Babasyan S, Freer H, Watts C, Rollins A, Osterrieder N, Wagner B. Deletion of the ORF2 gene of the neuropathogenic equine herpesvirus type 1 strain Ab4 reduces virulence while maintaining strong immunogenicity. BMC Vet Res 2018 Aug 22;14(1):245.
- Grandolfo E, Parisi A, Ricci A, Lorusso E, de Siena R, Trotta A, Buonavoglia D, Martella V, Corrente M. High mortality in foals associated with Salmonella enterica subsp. enterica Abortusequi infection in Italy. J Vet Diagn Invest 2018 May;30(3):483-485.
- Wagner B, Perkins G, Babasyan S, Freer H, Keggan A, Goodman LB, Glaser A, Torsteinsdóttir S, Svansson V, Björnsdóttir S. Neonatal Immunization with a Single IL-4/Antigen Dose Induces Increased Antibody Responses after Challenge Infection with Equine Herpesvirus Type 1 (EHV-1) at Weanling Age. PLoS One 2017;12(1):e0169072.
- Goodman LB, Anderson RR, Slater M, Ortenberg E, Renshaw RW, Chilson BD, Laverack MA, Beeby JS, Dubovi EJ, Glaser AL. High-throughput Detection of Respiratory Pathogens in Animal Specimens by Nanoscale PCR. J Vis Exp 2016 Nov 28;(117).
- Heerkens TM. Equine herpesvirus-1, non-neurogenic pathotype, in a 9-year-old American Saddlebred with neurological signs. Can Vet J 2009 Mar;50(3):297-300.
- Tallmadge RL, Laverack M, Lejeune M, Crossley B, Diel DG. A multiplex real-time PCR assay for detection of equid herpesvirus 1 and 4. Sci Rep 2025 Oct 31;15(1):38201.
- 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.
- Ali AAH, Abdallah F, Shemies OA, Kotb G, Nafea MR. Molecular characterization of equine herpes viruses type 1 and 4 among Arabian horse populations in Egypt during the period between 2021 and 2022. Open Vet J 2024 Jan;14(1):534-544.
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