title: Development and application of molecular methods (PCR) for detection of Tasmanian Atlantic salmon reovirus.
abstract: Molecular (PCR) diagnostic tests for the detection and identification of aquareovirus in general, and Tasmanian Atlantic salmon reovirus (TSRV) specifically, were developed, and their diagnostic sensitivity and specificity were determined and compared with virus isolation in cell culture. Intralaboratory and interlaboratory comparison of PCR (conventional hemi-nested RT-PCR & RT-qPCR) and virus isolation in cell culture using finfish cell lines, CHSE-214 and EPC, was carried out for the detection and identification of TSRV using field samples of farmed Atlantic salmon Salmo salar, L. from various aquaculture sites around Tasmania. The interlaboratory comparison of diagnostic methods was carried out between two laboratories, AAHL-CSIRO and DPIPWE-Tasmania. A total of 144 fish from nine sites (12-33 fish per site) were sampled from two regions of Tasmania (Tamar River estuary in the north and Huon River estuary in the south-east) during late spring to early summer of 2009, and the data were analysed using different statistical approaches. The prevalence of TSRV ranged from 6% to 22% in both regions. All the diagnostic methods (data from both laboratories) had high specificity, while the estimated sensitivity varied between tests with RT-qPCR being the most sensitive (95.2%) method followed by virus isolation and then conventional hemi-nested RT-PCR.
authors.affiliation: National Centre for Marine Conservation and Resource Sustainability, University of Tasmania, Launceston, TAS, Australia.
authors.forename: S C
authors.initials: SC
authors.lastname: Zainathan
authors.affiliation: Australian Animal Health Laboratory, CSIRO Animal, Food and Health Sciences, Geelong, VIC, Australia.
authors.forename: G
authors.initials: G
authors.lastname: Carlile
authors.affiliation: Department of Primary Industries, Water and Environment, Department, Launceston, TAS, Australia.
authors.forename: J
authors.initials: J
authors.lastname: Carson
authors.affiliation: Australian Animal Health Laboratory, CSIRO Animal, Food and Health Sciences, Geelong, VIC, Australia.
authors.forename: K A
authors.initials: KA
authors.lastname: McColl
authors.affiliation: Australian Animal Health Laboratory, CSIRO Animal, Food and Health Sciences, Geelong, VIC, Australia.
authors.forename: M St J
authors.initials: MS
authors.lastname: Crane
authors.affiliation: Australian Animal Health Laboratory, CSIRO Animal, Food and Health Sciences, Geelong, VIC, Australia.
authors.forename: L M
authors.initials: LM
authors.lastname: Williams
authors.affiliation: Australian Animal Health Laboratory, CSIRO Animal, Food and Health Sciences, Geelong, VIC, Australia.
authors.forename: J
authors.initials: J
authors.lastname: Hoad
authors.affiliation: Australian Animal Health Laboratory, CSIRO Animal, Food and Health Sciences, Geelong, VIC, Australia.
authors.forename: N J G
authors.initials: NJ
authors.lastname: Moody
authors.affiliation: Department of Environment and Heritage Protection, Department, Brisbane, QLD, Australia.
authors.forename: H M
authors.initials: HM
authors.lastname: Aiken
authors.affiliation: Veterinary Microbiology, The University of Melbourne, Parkville, VIC, Australia.
authors.forename: G F
authors.initials: GF
authors.lastname: Browning
authors.affiliation: National Centre for Marine Conservation and Resource Sustainability, University of Tasmania, Launceston, TAS, Australia.
authors.forename: B F
authors.initials: BF
authors.lastname: Nowak
mesh_terms: D000818:Animals; D017756:Aquaculture / Q000379:methods*; D002460:Cell Line; D005393:Fish Diseases / Q000175:diagnosis* / Q000453:epidemiology / Q000821:virology; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000662:veterinary*; D015995:Prevalence; D012087:Reoviridae / Q000235:genetics / Q000502:physiology*; D012088:Reoviridae Infections / Q000175:diagnosis / Q000453:epidemiology / Q000662:veterinary* / Q000821:virology; D020317:Salmo salar / Q000821:virology; D012680:Sensitivity and Specificity; D013648:Tasmania
keywords: RT-PCR; RT-qPCR; Salmo salar L.; Tasmania; Tasmanian Atlantic salmon reovirus; diagnosis