Detection of the co infection of different grass carp reovirus strains using dsRNA sequencing technology

Abstract:

Viral genomic dsRNAs were extracted from Ctenopharyngodon idellus kidney (CIK) cells inoculated by diseased grass carp visceral organs with hemorrhagic symptom, and analyzed by agarose gel to show the typical genomic pattern of Aquareovirus. The purified viral pathogen caused typical cytopathic effects in CIK cells. A simplified dsRNA sequencing technology modified from FLAC (full length amplification of cDNA) method was employed to sequencing the partial viral dsRNA genomic fragments. Four full-length dsRNA genomic fragments of the amplified cDNA fragments were successfully sequenced. Phylogenetic analysis was performed to identify two different viral strains in our sample, which indicated that there were two GCRV strains, named as JX01 and JX02 respectively, causing disease in grass carp in Jiangxi province. The data further suggested that we could monitor the pandemic viral strains using our simplified dsRNA sequencing technology. Our research thus provides a new technical method for the prevention, control and epidemiology of grass carp reovirus disease.