Detection and quantitation of equid gammaherpesviruses (EHV-2, EHV-5) in nasal swabs using an accredited standardised quantitative PCR method.
Abstract: Equid gammaherpesviruses-2 and -5 are involved in respiratory problems, with potential clinical manifestations such as nasal discharge, pharyngitis and swollen lymph nodes. These viruses are sometimes associated with a poor-performance syndrome, which may result in a significant and negative economic impact for the horse industry. The aim of the present study was to develop and validate quantitative PCR methods for the detection and quantitation of EHV-2 and EHV-5 in equine respiratory fluids. Two distinct tests were characterised: (a) for the qPCR alone and (b) for the whole method (extraction and qPCR) according to the standard model AFNOR XP U47-600-2 (viz., specificity, quantifiable sensibility, linearity, accuracy, range of application, trueness, precision, repeatability and precision of reproducibility). EHV-2 and EHV-5 detection were performed on nasal swabs collected from 172 horses, all of which exhibited clinical signs of respiratory disease. The data revealed a high rate of EHV-2/EHV-5 co-detection that was correlated significantly with age. Viral load of EHV-2 was significantly higher in young horses whereas viral load of EHV-5 was not significantly different with age.
Copyright © 2013 Elsevier B.V. All rights reserved.
Publication Date: 2013-12-24 PubMed ID: 24370678DOI: 10.1016/j.jviromet.2013.12.008Google Scholar: Lookup
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- Journal Article
- Research Support
- Non-U.S. Gov't
Summary
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This study focused on developing a set of quantitative PCR tests for the detection and quantification of Equid gammaherpesviruses-2 and -5 in horse respiratory fluids. These viruses are implicated in respiratory diseases in horses and can have economic implications for the horse industry.
Objective of the Study
- The main objective of this research was to create and validate quantitative PCR tests for detecting and measuring the levels of EHV-2 and EHV-5 in the respiratory fluids of horses.
Methods Used
- Two distinct tests were characterised in the study: one was for the qPCR method alone and the other included the entire method of extraction and qPCR.
- The study used the standard model AFNOR XP U47-600-2, which assessed a range of factors such as specificity, measurable sensitivity, linearity, precision, range of application, and so on.
Sampling
- Nasal swabs were collected from 172 horses, all exhibiting signs of respiratory disease, and were tested for EHV-2 and EHV-5.
Findings
- The results revealed a high rate of co-detection of EHV-2 and EHV-5, which was significantly correlated with age.
- It was found that the viral load of EHV-2 was significantly higher in young horses, while the viral load of EHV-5 did not show a significant difference with age.
Implications of the Study
- The high co-detection rate of EHV-2 and EHV-5, as well as the higher viral load of EHV-2 in young horses, signifies the potential for a high impact on horses’ health—particularly young ones—and consequently, on the economic aspects of the horse industry.
- This study can help in early detection and management of these viruses, potentially reducing the negative impact on the horse industry.
Cite This Article
APA
Hue ES, Fortier GD, Fortier CI, Leon AM, Richard EA, Legrand LJ, Pronost SL.
(2013).
Detection and quantitation of equid gammaherpesviruses (EHV-2, EHV-5) in nasal swabs using an accredited standardised quantitative PCR method.
J Virol Methods, 198, 18-25.
https://doi.org/10.1016/j.jviromet.2013.12.008 Publication
Researcher Affiliations
- Frank Duncombe Laboratory-LABÉO, 1 route de Rosel, 14053 Caen Cedex 4, France; Normandie Universite, 14000 Caen, France; Unite Risques Microbiens (U2RM), EA 4655, 14032 Caen, France; Hippolia Foundation, La Maison du cheval, 6 avenue du Maréchal Montgomery, 14000 Caen, France. Electronic address: Erika.Hue@calvados.fr.
- Frank Duncombe Laboratory-LABÉO, 1 route de Rosel, 14053 Caen Cedex 4, France; Normandie Universite, 14000 Caen, France; Unite Risques Microbiens (U2RM), EA 4655, 14032 Caen, France; Hippolia Foundation, La Maison du cheval, 6 avenue du Maréchal Montgomery, 14000 Caen, France. Electronic address: Guillaume.Fortier@calvados.fr.
- Frank Duncombe Laboratory-LABÉO, 1 route de Rosel, 14053 Caen Cedex 4, France; Hippolia Foundation, La Maison du cheval, 6 avenue du Maréchal Montgomery, 14000 Caen, France. Electronic address: Christine.Fortier@calvados.fr.
- Frank Duncombe Laboratory-LABÉO, 1 route de Rosel, 14053 Caen Cedex 4, France; Normandie Universite, 14000 Caen, France; Unite Risques Microbiens (U2RM), EA 4655, 14032 Caen, France; Hippolia Foundation, La Maison du cheval, 6 avenue du Maréchal Montgomery, 14000 Caen, France. Electronic address: Albertine.Leon@calvados.fr.
- Frank Duncombe Laboratory-LABÉO, 1 route de Rosel, 14053 Caen Cedex 4, France; Hippolia Foundation, La Maison du cheval, 6 avenue du Maréchal Montgomery, 14000 Caen, France. Electronic address: Eric.Richard@calvados.fr.
- Frank Duncombe Laboratory-LABÉO, 1 route de Rosel, 14053 Caen Cedex 4, France; Normandie Universite, 14000 Caen, France; Unite Risques Microbiens (U2RM), EA 4655, 14032 Caen, France; Hippolia Foundation, La Maison du cheval, 6 avenue du Maréchal Montgomery, 14000 Caen, France. Electronic address: Loic.Legrand@calvados.fr.
- Frank Duncombe Laboratory-LABÉO, 1 route de Rosel, 14053 Caen Cedex 4, France; Normandie Universite, 14000 Caen, France; Unite Risques Microbiens (U2RM), EA 4655, 14032 Caen, France; Hippolia Foundation, La Maison du cheval, 6 avenue du Maréchal Montgomery, 14000 Caen, France. Electronic address: Stephane.Pronost@calvados.fr.
MeSH Terms
- Animals
- Gammaherpesvirinae / genetics
- Herpesviridae Infections / virology
- Horse Diseases / virology
- Horses / virology
- Nose / virology
- Polymerase Chain Reaction / methods
- Reproducibility of Results
- Respiratory Tract Diseases / virology
- Viral Load / genetics
Citations
This article has been cited 18 times.- Stasiak K, Dunowska M, Rola J. Genetic Diversity of Equid Herpesvirus 5 in Temporal Samples from Mares and Their Foals at Three Polish National Studs. Int J Mol Sci 2025 Aug 27;26(17).
- James K, Chappell DE, Craig B, Pariseau C, Wright C, van Harreveld P, Barnum S, Pusterla N. Investigation of Selected Prevalence Factors Associated with EHV-2 and/or EHV-5 Infection in Horses with Acute Onset of Fever and Respiratory Signs. Viruses 2025 Apr 25;17(5).
- Cardeti G, Manna G, Cersini A, Nardini R, Rosati S, Reina R, Cittadini M, Sittinieri S, Altigeri A, Marcario GA, Scicluna MT. Horse Innate Immunity in the Control of Equine Infectious Anemia Virus Infection: A Preliminary Study. Viruses 2024 Nov 21;16(12).
- Charbonnel A, Lavoie JP, Juette T, St-Sauveur VG, Denis S, Gagnon CA, Leclère M. Comparison of nasal swabs and handmade foam cubes for detecting equine herpesvirus 5 (EHV-5) by quantitative polymerase chain reaction (qPCR). Can J Vet Res 2025 Jan;89(1):11-16.
- Badr C, Souiai O, Arbi M, El Behi I, Essaied MS, Khosrof I, Benkahla A, Chabchoub A, Ghram A. Epidemiological and Phylogeographic Study of Equid Herpesviruses in Tunisia. Pathogens 2022 Sep 5;11(9).
- Onasanya AE, El-Hage C, Diaz-Méndez A, Vaz PK, Legione AR, Browning GF, Devlin JM, Hartley CA. Whole genome sequence analysis of equid gammaherpesvirus -2 field isolates reveals high levels of genomic diversity and recombination. BMC Genomics 2022 Aug 30;23(1):622.
- 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).
- Temesgen T, Getachew Y, Negussie H. Molecular Identification of Equine Herpesvirus 1, 2, and 5 in Equids with Signs of Respiratory Disease in Central Ethiopia. Vet Med (Auckl) 2021;12:337-345.
- 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).
- Stasiak K, Dunowska M, Trewick S, Rola J. Genetic Variation in the Glycoprotein B Sequence of Equid Herpesvirus 5 among Horses of Various Breeds at Polish National Studs. Pathogens 2021 Mar 9;10(3).
- Bond SL, Workentine M, Hundt J, Gilkerson JR, Léguillette R. Effects of nebulized dexamethasone on the respiratory microbiota and mycobiota and relative equine herpesvirus-1, 2, 4, 5 in an equine model of asthma. J Vet Intern Med 2020 Jan;34(1):307-321.
- 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.
- Azab W, Bedair S, Abdelgawad A, Eschke K, Farag GK, Abdel-Raheim A, Greenwood AD, Osterrieder N, Ali AAH. Detection of equid herpesviruses among different Arabian horse populations in Egypt. Vet Med Sci 2019 Aug;5(3):361-371.
- 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.
- Doubli-Bounoua N, Richard EA, Léon A, Pitel PH, Pronost S, Fortier G. Multiple molecular detection of respiratory viruses and associated signs of airway inflammation in racehorses. Virol J 2016 Nov 29;13(1):197.
- Hue ES, Fortier CI, Laurent AM, Quesnelle YF, Fortier GD, Legrand LJ, Pronost SL. Development and Validation of a Quantitative PCR Method for Equid Herpesvirus-2 Diagnostics in Respiratory Fluids. J Vis Exp 2016 Mar 17;(109).
- Couëtil LL, Cardwell JM, Gerber V, Lavoie JP, Léguillette R, Richard EA. Inflammatory Airway Disease of Horses--Revised Consensus Statement. J Vet Intern Med 2016 Mar-Apr;30(2):503-15.
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