title: Use of automated real-time reverse transcription-polymerase chain reaction (RT-PCR) to monitor experimental swine vesicular disease virus infection in pigs.
abstract: Automated real-time RT-PCR was evaluated as a diagnostic tool for swine vesicular disease virus (SVDV) infection on a range of samples (vesicular epithelium, serum, nasal swabs, faeces) from four inoculated and three in-contact pigs over a period of 28 days. Traditional diagnostic procedures (virus isolation, and ELISAs for antigen and antibody) were used in parallel. Each inoculated pig developed a significant viraemia and clinical disease, and excreted virus, which was transmitted to the in-contact animals. The latter, however, developed only a short-lived, low-level viraemia and no clinical disease. The RT-PCR and virus isolation were generally comparable in detecting SVDV in the serum and nasal swabs from inoculated and in-contact pigs up to day 6 after infection; it was possible, however, to isolate virus for a longer period from the faeces of a few pigs. This suggested that further optimization of the template extraction method was required to counteract the effects of RT-PCR inhibitors in faeces. It was concluded that the automated real-time RT-PCR is a useful diagnostic method for SVD in clinically or subclinically affected pigs and contributed to the study of the pathogenesis of SVD in the pigs.
authors.affiliation: Institute for Animal Health, Pirbright Laboratory, Ash Road, Pirbright, Woking, Surrey GU24 0NF, UK.
authors.forename: S M
authors.initials: SM
authors.lastname: Reid
authors.forename: D J
authors.initials: DJ
authors.lastname: Paton
authors.forename: G
authors.initials: G
authors.lastname: Wilsden
authors.forename: G H
authors.initials: GH
authors.lastname: Hutchings
authors.forename: D P
authors.initials: DP
authors.lastname: King
authors.forename: N P
authors.initials: NP
authors.lastname: Ferris
authors.forename: S
authors.initials: S
authors.lastname: Alexandersen
mesh_terms: D000818:Animals; D018562:Disease Transmission, Infectious; D029822:Enterovirus B, Human / Q000235:genetics / Q000276:immunology / Q000472:pathogenicity*; D004797:Enzyme-Linked Immunosorbent Assay / Q000662:veterinary; D012367:RNA, Viral / Q000032:analysis; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods / Q000662:veterinary*; D013552:Swine; D013553:Swine Diseases / Q000175:diagnosis* / Q000821:virology; D013555:Swine Vesicular Disease / Q000473:pathology / Q000635:transmission / Q000821:virology
chemical_list: D012367:RNA, Viral