Prevalence of equine gammaherpesviruses on breeding farms in Turkey and development of a TaqMan MGB real-time PCR to detect equine herpesvirus 5 (EHV-5).
Abstract: Equine herpesvirus type 2 (EHV-2) and EHV-5 are members of the subfamily Gammaherpesvirinae. The viruses are detected in horses with upper respiratory tract disease and are associated with low performance in racehorses. The aim of the current study was to use nested PCR to investigate the epidemiology of EHV-2 and EHV-5 in Arabian horse populations from breeding farms located in three different cities (Eskişehir, Malatya, and Bursa) in Turkey, using a real-time quantitative PCR (qPCR) with a TaqMan® minor-groove-binder (MGB) probe to detect EHV-5. Screening of blood and ocular and nasal swab samples by nested PCR showed the prevalence of EHV-2 and EHV-5 to be 59 % and 62 %, respectively, with a coinfection rate of 45 %. Thirty-seven isolates from blood samples were identified as EHV-2 using nested PCR. To develop the EHV-5 qPCR, a pair of primers and an MGB probe were designed based on a highly conserved genomic region encoding glycoprotein B (gB). The detection limit of the qPCR was 10 molecules per reaction, and it specifically detected EHV-5 and no other herpesviruses infecting horses (EHV-1, EHV-2, or EHV-4). When applied to field samples, the assay proved to be more sensitive than a well-established nested PCR. Therefore, the qPCR developed in this study provides a rapid, reliable, and sensitive diagnostic assay for the detection of EHV-5, and it complements other diagnostic procedures for equine respiratory disease.
Publication Date: 2014-07-10 PubMed ID: 25008897DOI: 10.1007/s00705-014-2165-5Google Scholar: Lookup
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- Evaluation Study
- Journal Article
- Research Support
- Non-U.S. Gov't
Summary
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The research article reveals the high prevalence of Equine herpesvirus type 2 (EHV-2) and EHV-5 in Arabian horse populations in Turkey. A new diagnostic technique involving real-time PCR was also developed, proving more accurate and efficient in detecting EHV-5.
Research Objective
- The primary goal of the study was to investigate the prevalence of EHV-2 and EHV-5 in Arabian horse populations from breeding farms in Turkey. This investigation utilized nested PCR to understand the epidemiology of the viruses, which are associated with respiratory tract infections and low performance in racehorses.
- Another central goal was the development of a real-time quantitative PCR (qPCR) technique with a TaqMan® minor-groove-binder (MGB) probe to offer speedy and reliable detection of EHV-5.
Methodology
- Blood, and ocular and nasal swab samples were taken from the horses for screening by nested PCR. The research was conducted in three different cities (Eskişehir, Malatya, and Bursa).
- To create the EHV-5 qPCR, a set of primers and an MGB probe were designed. This equipment relies on the highly-conserved genomic region that encodes glycoprotein B (gB).
Key Findings
- The prevalence of EHV-2 and EHV-5 was found to be 59% and 62%, respectively, while the coinfection rate was around 45%. Thirty-seven isolates were identified as EHV-2 using nested PCR.
- The qPCR assay developed was effective in detecting EHV-5 down to a level of 10 molecules per reaction. It was specific to EHV-5 and didn’t respond to other types of herpesviruses found in horses (EHV-1, EHV-2, or EHV-4).
- When applied to field samples, the qPCR assay demonstrated a superior sensitivity compared to the established nested PCR technique, making it a promising tool for the swift and accurate detection of EHV-5.
Conclusions
- The research highlights the high prevalence of EHV-2 and EHV-5 in Arabian horse populations across Turkey, indicating the critical need for effective detection methods to manage and prevent viral outbreaks.
- The newly developed qPCR assay in this study represents a substantial improvement over existing techniques, presenting a faster, more reliable, and more sensitive method for the detection of EHV-5. Therefore, it is an excellent addition to other diagnostic methods for equine respiratory disease.
Cite This Article
APA
Akkutay AZ, Osterrieder N, Damiani A, Tischer BK, Borchers K, Alkan F.
(2014).
Prevalence of equine gammaherpesviruses on breeding farms in Turkey and development of a TaqMan MGB real-time PCR to detect equine herpesvirus 5 (EHV-5).
Arch Virol, 159(11), 2989-2995.
https://doi.org/10.1007/s00705-014-2165-5 Publication
Researcher Affiliations
- Department of Virology, Faculty of Veterinary Medicine, Ankara University, Diskapi, 06110, Ankara, Turkey, akkutay@ankara.edu.tr.
MeSH Terms
- Animals
- Breeding
- DNA, Viral / genetics
- Herpesviridae Infections / diagnosis
- Herpesviridae Infections / epidemiology
- Herpesviridae Infections / veterinary
- Herpesviridae Infections / virology
- Herpesvirus 1, Equid / genetics
- Herpesvirus 1, Equid / isolation & purification
- Horse Diseases / diagnosis
- Horse Diseases / epidemiology
- Horse Diseases / virology
- Horses
- Real-Time Polymerase Chain Reaction / methods
- Taq Polymerase / metabolism
- Turkey / epidemiology
Citations
This article has been cited 9 times.- Stasiak K, Dunowska M, Rola J. Kinetics of the Equid Herpesvirus 2 and 5 Infections among Mares and Foals from Three Polish National Studs.. Viruses 2022 Mar 29;14(4).
- Scheurer L, Bachofen C, Hardmeier I, Lechmann J, Schoster A. Prevalence of Nasal Shedding of Equid Gammaherpesviruses in Healthy Swiss Horses.. Viruses 2021 Aug 25;13(9).
- Chen K, Kong M, Liu J, Jiao J, Zeng Z, Shi L, Bu X, Yan Y, Chen Y, Gao R, Liu X, Wang X, Hu J, Hu S, Jiao X, Liu X, Gu M. Rapid differential detection of subtype H1 and H3 swine influenza viruses using a TaqMan-MGB-based duplex one-step real-time RT-PCR assay.. Arch Virol 2021 Aug;166(8):2217-2224.
- Zarski LM, Giessler KS, Jacob SI, Weber PSD, McCauley AG, Lee Y, Soboll Hussey G. Identification of Host Factors Associated with the Development of Equine Herpesvirus Myeloencephalopathy by Transcriptomic Analysis of Peripheral Blood Mononuclear Cells from Horses.. Viruses 2021 Feb 24;13(3).
- Zarski LM, Weber PSD, Lee Y, Soboll Hussey G. Transcriptomic Profiling of Equine and Viral Genes in Peripheral Blood Mononuclear Cells in Horses during Equine Herpesvirus 1 Infection.. Pathogens 2021 Jan 7;10(1).
- Van Cleemput J, Poelaert KCK, Laval K, Nauwynck HJ. Unravelling the first key steps in equine herpesvirus type 5 (EHV5) pathogenesis using ex vivo and in vitro equine models.. Vet Res 2019 Feb 18;50(1):13.
- Marenzoni ML, Stefanetti V, Danzetta ML, Timoney PJ. Gammaherpesvirus infections in equids: a review.. Vet Med (Auckl) 2015;6:91-101.
- Stasiak K, Dunowska M, Rola J. Prevalence and sequence analysis of equid herpesviruses from the respiratory tract of Polish horses.. Virol J 2018 Jul 11;15(1):106.
- Zhang Z, Liu D, Sun W, Liu J, He L, Hu J, Gu M, Wang X, Liu X, Hu S, Chen S, Peng D, Liu X. Multiplex one-step Real-time PCR by Taqman-MGB method for rapid detection of pan and H5 subtype avian influenza viruses.. PLoS One 2017;12(6):e0178634.
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