title: An RT-PCR-based method for the diagnosis of the sleeping disease virus in experimentally and naturally infected salmonids.
abstract: The sleeping disease (SD) of rainbow trout (Oncorhynchus mykiss) is a worldwide disease for which the causative agent, the sleeping disease virus (SDV), has been recently characterized as an atypical alphavirus (Villoing et al. 2000). Up to now, no diagnostic tools were available and thus no epidemiological studies have been undertaken to evaluate the occurrence of this disease on the field. We present in this paper a sensitive and highly specific 1 working day method, which allows the detection of SDV from experimentally and naturally infected fishes. This method, based on a reverse transcriptase/polymerase chain reaction (RT-PCR) assay on total RNA extracted from SDV-infected fish organs, enables the specific DNA amplification of part of the gene encoding the SDV glycoprotein E2, as early as 2 d post-infection (d.p.i.) and as late as 70 d.p.i., at which time clinical signs of infection are no longer apparent. Moreover, we show that this RT-PCR method can be successfully used for the diagnosis of fish infected by a closely related virus, namely salmon pancreas disease virus (SPDV). This report is the first description of a very powerful diagnostic assay which could provide a more accurate replacement for the classical virological, histological and immunochemistry methods.
authors.affiliation: Unité Virologie et Immunologie moléculaires, Institut National de la Recherche Agronomique, INRA, Jouy-en-Josas, France.
authors.forename: S
authors.initials: S
authors.lastname: Villoing
authors.forename: J
authors.initials: J
authors.lastname: Castric
authors.forename: J
authors.initials: J
authors.lastname: Jeffroy
authors.forename: A
authors.initials: A
authors.lastname: Le Ven
authors.forename: R
authors.initials: R
authors.lastname: Thiery
authors.forename: M
authors.initials: M
authors.lastname: Bremont
mesh_terms: D000524:Alphavirus; D018354:Alphavirus Infections / Q000175:diagnosis / Q000662:veterinary*; D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D005393:Fish Diseases / Q000175:diagnosis*; D008969:Molecular Sequence Data; D017686:Oncorhynchus mykiss / Q000821:virology*; D010182:Pancreatic Diseases / Q000175:diagnosis / Q000662:veterinary; D016133:Polymerase Chain Reaction / Q000662:veterinary*; D014759:Viral Envelope Proteins / Q000737:chemistry / Q000235:genetics
chemical_list: D014759:Viral Envelope Proteins