A GENETIC PROBE FOR IDENTIFICATION OF THE TURBOT AQUAREOVIRUS IN INFECTED CELL-CULTURES

A nucleic acid hybridization assay has been developed that can detect small amounts of the turbot aquareovirus (TRV) in infected cells. Comp lementary DNAs were synthesized from the dsRNA genome segments of TRV and used to generate a large number of recombinant plasmids. Plasmids corresponding to genomic segments 1 through 8 were identified by North ern blot analysis. A randomly selected clone, Clone 66, hybridizing to genome Segment 8, showed high specificity hybridizing only RNA from T RV, and not with the RNAs of 4 other aquareoviruses, or with RNA extra cted from infectious pancreatic necrosis virus (IPNV), infectious hema topoietic necrosis virus (IHNV) or with uninfected cells. A P-32-label ed probe was able to detect as little as 50 ng of TRV purified virus d sRNA. Time course experiments indicated that TRV RNA can be detected i n infected cells 96 h post-inoculation, a time when cytopathic effect is still not evident. These results indicate that the dot blot assay d escribed here could be used as an effective diagnostic tool for the de tection of TRV infections.