title: 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.
authors.affiliation: 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.
authors.forename: Erika S
authors.initials: ES
authors.lastname: Hue
authors.affiliation: 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.
authors.forename: Guillaume D
authors.initials: GD
authors.lastname: Fortier
authors.affiliation: 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.
authors.forename: Christine I
authors.initials: CI
authors.lastname: Fortier
authors.affiliation: 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.
authors.forename: Albertine M
authors.initials: AM
authors.lastname: Leon
authors.affiliation: 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.
authors.forename: Eric A
authors.initials: EA
authors.lastname: Richard
authors.affiliation: 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.
authors.forename: Loïc J
authors.initials: LJ
authors.lastname: Legrand
authors.affiliation: 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.
authors.forename: Stéphane L
authors.initials: SL
authors.lastname: Pronost
mesh_terms: D000818:Animals; D018142:Gammaherpesvirinae / Q000235:genetics*; D006566:Herpesviridae Infections / Q000821:virology*; D006734:Horse Diseases / Q000821:virology; D006736:Horses / Q000821:virology*; D009666:Nose / Q000821:virology*; D016133:Polymerase Chain Reaction / Q000379:methods*; D015203:Reproducibility of Results; D012140:Respiratory Tract Diseases / Q000821:virology*; D019562:Viral Load / Q000235:genetics
keywords: Accredited standardised method; EHV-2; EHV-5; Horses; qPCR