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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 explore the isolation and identification of pathogens in cases of suspected fever with thrombocytopenia syndrome and to understand the characteristics of virus growth. Methods Vero E6 was used to isolate fever and thrombocytopenia syndrome Bunyavirus (SFTSV) from patients with anticoagulant blood samples. The virus was identified by serological and morphological methods. The isolated virus was inoculated In different cells, the changes of viral load in the supernatant of cells at different times were detected by real-time PCR. Results SFTSV was isolated from the anticoagulant samples of patients. Immunofluorescence showed that the virus-infected cell cultures could react positively with the serum of patients. Bunyavirus-like particles were observed under transmission electron microscope. SFTSV can infect Vero E6, Hep G2, THP-1 and other cells, but the virus replication in different cells significantly different levels. Among them, Vero E6 cells are more susceptible to SFTSV and the virus can proliferate to 4.89 × 109 copies / ml. While Hep-2 and HT29 cells can not be infected with SFTSV. Conclusion SFTSV was successfully isolated from blood samples of suspected fever with thrombocytopenia syndrome. The virus has a broader cell tropism, more susceptible to kidney-derived cells. These studies will provide important clues for the follow-up study of SFTSV related basic research.