Isolation and identification ofTupaiaorthoreovirus

Pathogenic viruses can harm acutely the life and health of laboratory tree shrews acutely; however, few papers exist regarding natural pathogenic virus infection in this species. Six fecal samples obtained from dead tree shrews were collected. The fecal supernatant infected Vero cell line resulted in cytopathic effects (CPE) after 72 h. The CPE included granulating, shrinking, rounding, seining and falling off. Electron microscopy showed the isolation was spherical, double-layered capsid, and about 75 nm in diameter. The purified isolation genome was 10 segments in a typical 3:3:4 arrangements, as shown by polyacrylamide gel electrophoresis (PAGE). The isolation was confirmed by RT-PCR assays targeting the conserved region of the L1 gene, sequence analysis and reconstruction of a phylogenetic tree. The isolation was a Tupaia Orthoreovirus (TRV), belonging to Mammalian Orthoreovirus (MRV). The obtained strain had the closest phylogenetic relationship to the MRV strain T3/Bat/Germany/342/08. As a zoonotic virus, the novel TRV strain was first isolated from wild tree shrews, which is significant for promoting tree shrew standardization and providing scientific data for preventing zoonotic tree shrew-to-human transmission.致病性病毒严重危害树鼩 (tree shrew, Tupaia)生命健康, 但却鲜见有关树鼩自然感染病毒的报道。该研究采集6份因腹泻死亡的野生树鼩粪便, 用Vero细胞进行病毒分离, 72 h后细胞发生病变, 特征为颗粒增多、破碎、变圆固缩、拉网及脱落等;电镜观察显示该病毒为球形, 双层衣壳, 完整直径~75 nm。纯化病毒经核酸PAGE电泳后呈现10个核酸节段, 并为典型的3:3:4排列。哺乳动物呼肠孤病毒(mammalian orthoreovirus, MRV) L1基因保守区RT-PCR检测、序列分析及进化树构建结果表明, 该病毒株属于MRV, 且与来源于蝙蝠的呼肠孤病毒分离株 (T3/Bat/Germany/342/08)同源性最高。MRV是人兽共患病病毒, 该实验首次从树鼩体内成功分离到MRV, 对今后研究制定实验树鼩病毒学控制标准具有指导意义, 同时为有效预防该病毒在树鼩与人类之间传播提供了实验依据。