云南省中山病毒的分离鉴定

Isolation and identication of Chuzan virus from Yunnan Province

为掌握云南省虫媒病毒的流行情况及基因分型,本试验在云南省师宗、江城和芒市设立监控点,对哨兵动物定期采血来监测其血清转阳情况,将阳性样品在BHK上传代以分离病毒,用RT-PCR及MNT鉴定病毒,通过特异性引物扩增分离毒株的Segment2(Seg-2)及Segment7(Seg-7)基因片段来分析其遗传特性。结果显示,在2014年从师宗监控点分离得到4株帕利亚姆病毒(PALV),经鉴定均为中山病毒(Chuzan virus,CHUV),并掌握了分离株Seg-2及Seg-7基因片段的遗传特征。本试验确认了CHUV在云南省的分离,为进一步掌握CHUV在中国的流行病学、致病性以及疫苗研究奠定良好基础。

In order to investigate the distribution and prevalence of arbovirus in Yunnan Province,three monitoring points were set at Shizong,Jiangcheng and Mangshi,blood samples of the sentinel animals were regularly collected and virus isolation were carried out through inoculating BHK cells,isolated strains were identified by RT-PCR amplification and neutralization assay,specific primers were designed to amplify the ORF regions of Segment2(Seg-2)and Segment7(Seg-7)of isolated strains,then cloned and sequenced.This study confirmed the isolation of Chuzan virus(CHUV)strain in Yunnan Province,and elucidated the evolutionary characteristics of the virus Seg-2 and Seg-7 genes.The results laid a foundation for further research on epidemiology and pathogenicity and vaccine of Chinese CHUV.