title: Duplex Real-Time RT-PCR Assays for the Detection and Typing of Epizootic Haemorrhagic Disease Virus.
abstract: Epizootic haemorrhagic disease virus (EHDV) may cause severe clinical episodes in some species of deer and sometimes in cattle. Laboratory diagnosis provides a basis for the design and timely implementation of disease control measures. There are seven distinct EHDV serotypes, VP2 coding segment 2 being the target for serotype specificity. This paper reports the development and validation of eight duplex real-time RT-PCR assays to simultaneously amplify the EHDV target (S9 for the pan-EHDV real-time RT-PCR assay and S2 for the serotyping assays) and endogenous control gene Beta-actin. Analytical and diagnostic sensitivity and specificity, inter- and intra-assay variation and efficiency were evaluated for each assay. All were shown to be highly specific and sensitive.
authors.affiliation: ANSES/INRA/ENVA-UPEC, UMR 1161 Virologie, 23 avenue du général de Gaulle-94700 Maisons Alfort-France.
authors.forename: Cyril
authors.initials: C
authors.lastname: Viarouge
authors.affiliation: ANSES/INRA/ENVA-UPEC, UMR 1161 Virologie, 23 avenue du général de Gaulle-94700 Maisons Alfort-France.
authors.forename: Emmanuel
authors.initials: E
authors.lastname: Breard
authors.affiliation: ANSES/INRA/ENVA-UPEC, UMR 1161 Virologie, 23 avenue du général de Gaulle-94700 Maisons Alfort-France.
authors.forename: Stephan
authors.initials: S
authors.lastname: Zientara
authors.affiliation: ANSES/INRA/ENVA-UPEC, UMR 1161 Virologie, 23 avenue du général de Gaulle-94700 Maisons Alfort-France.
authors.forename: Damien
authors.initials: D
authors.lastname: Vitour
authors.affiliation: ANSES/INRA/ENVA-UPEC, UMR 1161 Virologie, 23 avenue du général de Gaulle-94700 Maisons Alfort-France.
authors.forename: Corinne
authors.initials: C
authors.lastname: Sailleau
mesh_terms: D000818:Animals; D002417:Cattle; D017931:DNA Primers / Q000378:metabolism; D015342:DNA Probes / Q000378:metabolism; D018164:Hemorrhagic Disease Virus, Epizootic / Q000145:classification* / Q000235:genetics*; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*; D015203:Reproducibility of Results; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D065288:Serogroup
chemical_list: D017931:DNA Primers; D015342:DNA Probes