title: Real-time RT-PCR for detection of equine influenza and evaluation using samples from horses infected with A/equine/Sydney/2007 (H3N8).
abstract: Equine influenza (EI) virus (H3N8) was identified in the Australian horse population for the first time in August 2007. The principal molecular diagnostic tool used for detection was a TaqMan real-time reverse transcription-polymerase chain reactions (RT-PCR) assay specific for the matrix (MA) gene of influenza virus type A (IVA). As this assay is not specific for EI, we developed a new EI H3-specific TaqMan assay targeting the haemagglutinin (HA) gene of all recent EI H3 strains. The IVA and the EI H3 TaqMan assays were assessed using in vitro transcribed RNA template, virus culture, diagnostic samples from the outbreak and samples from experimentally infected horses. The EI H3 TaqMan assay had a higher diagnostic sensitivity (DSe) when compared to the IVA TaqMan assay and also when using a conventional PCR for EI H3 as a standard of comparison. The performance of both TaqMan assays was compared with an antigen detection ELISA and virus isolation using nasal swabs collected daily from horses experimentally infected with the outbreak strain A/equine/Sydney/2888-8/2007. The EI H3 TaqMan assay was the most sensitive of the assays, able to detect EI from day 1 or 2 post-challenge, as early as virus isolation, and before clinical signs of disease were observed.
authors.affiliation: CSIRO Livestock Industries, Australian Animal Health Laboratory, 5 Portarlington Road (Private Bag 24), Geelong, VIC 3220, Australia.
authors.forename: Adam J
authors.initials: AJ
authors.lastname: Foord
authors.forename: Paul
authors.initials: P
authors.lastname: Selleck
authors.forename: Axel
authors.initials: A
authors.lastname: Colling
authors.forename: Jessica
authors.initials: J
authors.lastname: Klippel
authors.forename: Deborah
authors.initials: D
authors.lastname: Middleton
authors.forename: Hans G
authors.initials: HG
authors.lastname: Heine
mesh_terms: D000818:Animals; D001483:Base Sequence; D005814:Genes, Viral; D006734:Horse Diseases / Q000175:diagnosis / Q000821:virology*; D006736:Horses; D053123:Influenza A Virus, H3N8 Subtype / Q000235:genetics / Q000302:isolation & purification*; D009976:Orthomyxoviridae Infections / Q000175:diagnosis / Q000662:veterinary* / Q000821:virology; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods / Q000662:veterinary*; D012680:Sensitivity and Specificity; D054334:Viral Regulatory and Accessory Proteins
chemical_list: D054334:Viral Regulatory and Accessory Proteins