Isolation and identification of Getah virus from Culicoides in Yunnan

Objective To discuss the taxonomy and molecular characteristics of one virus strain (SZC30) isolated from Culicoides in Yunnan. Methods Culicoides were collected with light trap method in the Wulong Village of Shizong County of Yunnan Province in July, 2013. BHK-21 and C6/36 cells were used for virus isolation. The positive isolates were inoculated into brain of one-day suckling mice. Alphavirus and Getah virus specific primers were used to amplify the genome of the virus isolation by RT-PCR. The products of RT-PCR were sequenced. Clustal X1.83, DNAStar, Mega5.1 were used for bioinformatics analysis. Results Totally 3 500 culicoides were collected and divided into 41 batches for virus isolation. One isolate (SZC30) produced cytopathic effect (CPE) on BHK-21 and C6/36 cells; the result of RT-PCR with Alphavirus and Getah virus NS1 specific primer were positive; the sequence analysis of NS1 gene suggested that SZC30 and two Getah virus strains (YN0540, SC1210) from China were in the same evolutionary branching, the nucleotide homology were 97%-100%, and the amino acid was 97%-100%. Conclusions SZC30 isolated in Yunnan province was identified as Getah virus. Key words: Getah virus; Culicoides; Isolation; Identification