一株发热伴血小板减少综合征布尼亚病毒的分离鉴定及其生长特性研究

Study of isolation and identification of severe fever with thrombocytopenia syndrome bunyavirus and its growth characteristics

目的探索疑似发热伴血小板减少综合征病例中病原体的分离鉴定,了解病毒生长特性。方法利用非洲绿猴肾细胞Vero E6从患者抗凝血标本中分离发热伴血小板减少综合征布尼亚病毒(SFTSV),并通过血清学、形态学等方法对病毒进行鉴定;将分离的病毒接种不同细胞,利用荧光定量PCR检测不同时间细胞上清中病毒载量的变化。结果从患者抗凝血标本中分离出SFTSV;免疫荧光显示感染病毒的细胞培养物能与患者血清发生阳性反应;在透射电镜下能观察到布尼亚病毒样颗粒。SFTSV可感染Vero E6、Hep G2、THP-1等多种细胞,但病毒在不同细胞中的复制水平差异明显。其中Vero E6细胞对SFTSV较易感,病毒可增殖至4.89×109copy/ml。而Hep-2和HT29细胞不能被SFTSV感染。结论从疑似发热伴血小板减少综合征病例血液标本中成功分离出SFTSV。该病毒具有较广泛的细胞嗜性,对肾来源的细胞更易感。这些研究将为后续研究SFTSV的相关基础研究提供重要的线索。

Objective To isolate and identify the pathogen of one case of severe fever with thrombocytopenia syndrome,and also understand the growth characteristics of the pathogen. Methods We used Vero E6 cells to isolate severe fever with thrombocytopenia syndrome bunyavirus（ SFTSV） from the anticoagulant blood of the case,and subsequent virus identification was conducted with serological and morphological methods; the virus isolated was inoculated with different cells and the viral loads were detected at different times by fluorescence quantitative PCR. Results SFTSV was isolated from the anticoagulant blood. The immune fluorescence results showed that the reaction between the patient sera and the cell suspension was positive.We could observe the bunyavirus-like particles under transmission electron microscope. SFTSV could infect Vero E6,Hep G2 and THP- 1 cells,but the virus replication levels varied in these cells. Among them,Vero E6 cells were relatively susceptible to SFTSV,and the viral loads could reach 4. 89 × 109 copy / ml. SFTSV could not infect Hep- 2 and HT29 cells. Conclusion SFTSV was successfully isolated from the patient. SFTSV has a wide range of tissue and cell tropism. The cells from kidney organs are more likely to be infected. This study provides important evidence for the future study of SFTSV.