发热伴血小板减少综合征病毒在Vero细胞中的培养特性观察  

Observation on the culture characteristics of SFTSV in Vero cells

作　　者：胡逢蛟[1] 焦素黎[1] 王忠发[2] 倪红霞[1] 易波[1] 李永东[1] 

机构地区：[1]宁波市疾病预防控制中心,浙江宁波315010 [2]舟山市疾病预防控制中心,浙江舟山316000

出　　处：《中国卫生检验杂志》2014年第21期3070-3072,共3页Chinese Journal of Health Laboratory Technology

基　　金：2013年浙江省肾综合征出血热疫苗研究重点实验室开放课题(201301)

摘　　要：目的了解发热伴血小板减少综合征布尼亚病毒（SFTSV）在绿猴肾细胞（Vero细胞）中培养的特征。方法收集荧光定量逆转录聚合酶链式反应（荧光定量RT-PCR）阳性的发热伴血小板减少综合征（SFTS）阳性病人血清,应用Vero细胞对SFTSV进行分离培养。结果 24份核酸阳性病人标本有21份在Vero细胞中出现了细胞病变（CPE）,其中第1代5份、第2代8份及第3代5份,另有3份在培养到第6代~7代时才出现CPE。病毒分离率达87.5%（21/24）。并且毒株经Vero细胞连续传代后CPE出现的时间及进展均明显加快,一般均可稳定在5 d内出现明显的CPE。结论虽然SFTSV的初代培养周期较长（约14 d）,但经传代后CPE出现时间明显缩短,并且TCID50也较高。因此,应用Vero细胞分离SFTSV既直观又敏感,同时又是疫苗研究中应用最广泛的细胞基质。Objective To understand the cultured characteristics of Severe fever with thrombocytopenia syndrome bunyavirus （SFTSV） in the African green monkey kidney（Vero） cells. Methods The positive samples of the cases with Severe fever with ihrombocytopenia syndrome （SFIS）, which were identified by real -time PCR , were collected, and then inoculated into Vero cells for sFrsv !solating. Results 21 out of 24 serum, which were nucleic acid positive identified by real - time PCR, caused cytopathic effect （CPE） in Veto cells. Among them, 5 appeared in the first generation, 8 appeared in the second generation and 5 appeared in the third generation, and others were up to 6 ~ 7 generations to caused CPE appeared. The isolating rate was up to 87.5%. And the time of CPE appeared significantly speeded up by continuous passage culture. Generally CPE can be obviously observed stably in 5 d. Conclusion Although the primary culture period of SFTSV was longer than some viruses, CPE appeared time was significantly shortened after continuous passage culture, and the TCID50 titer was at the high level. Therefore, the advantages of isolating SFTSV with Vero ceils were intuitive and sensitive, and this cell lines was the commom paltform for vaccine development and research.

关 键 词：发热伴血小板减少综合征布尼亚病毒 细胞培养 细胞病变 

分 类 号：R181.36[医药卫生—流行病学]