In JoVE (1)
Other Publications (1)
Articles by Erika S. Hue in JoVE
Development and Validation of a Quantitative PCR Method for Equid Herpesvirus-2 Diagnostics in Respiratory Fluids Erika S. Hue1,2,3, Christine I. Fortier1,3, Aurélie M. Laurent1, Yann F. Quesnelle1, Guillaume D. Fortier1,2,3, Loïc J. Legrand1,2,3, Stephane L. Pronost1,2,3 1LABÉO Frank Duncombe, 2Unité de Risques Microbiens (U2RM, EA 4655), Normandy University, 3Hippolia Foundation Here, we present a protocol for the development and validation of a quantitative PCR method used for the detection and quantification of EHV-2 DNA in equine respiratory fluids. The EHV-2 qRT-PCR validation protocol involves a three-part procedure: development, characterization of qRT-PCR assay alone, and characterization of the whole analytical method.
Other articles by Erika S. Hue on PubMed
Detection and Quantitation of Equid Gammaherpesviruses (EHV-2, EHV-5) in Nasal Swabs Using an Accredited Standardised Quantitative PCR Method Journal of Virological Methods. Mar, 2014 | Pubmed ID: 24370678 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.