登革病毒感染C6/36、Vero、BHK-21和THP-1细胞CPE的比较及病毒检测  

CPE caused by dengue virus in C6/36,Vero,BHK-21 and THP 1 cells and virus test

作　　者：杨佳佳[1] 姜黎明[1] 罗佳[1] 叶超[1] 文送娇 孙强明[1] 

机构地区：[1]中国医学科学院北京协和医学院医学生物学研究所云南省重大传染病疫苗研发重点实验室,云南昆明650118

出　　处：《中国生物制品学杂志》2017年第4期362-365,371,共5页Chinese Journal of Biologicals

基　　金：国家自然科学基金面上项目(81171946);国家"十二五""重大新药创新"科技重大专项(2012ZX09104-302);云南省科技厅社会发展科技计划项目(2011CA016);云南自然科学基金项目(2012FB188;2016FA029);医学生物学研究所研究生创新基金(2015-018-004)

摘　　要：目的比较感染登革病毒(dengue virus,DV)的C6/36、Vero、BHK-21和THP-1细胞的CPE情况,并进行病毒检测。方法将分离自云南西双版纳的Ⅲ型DV中国株D13053和广东省疾病预防控制中心惠赠的Ⅱ型中国株D10156,以5 MOI感染C6/36、Vero、BHK-21和THP-1细胞,同时将Vero和C6/36细胞来源的DV,分别以5 MOI感染C6/36细胞,连续观察CPE至第10天,并利用DV通用引物,RT-PCR法鉴定细胞培养上清中的病毒。结果C6/36细胞对DV最敏感并有明显的病变,细胞几乎100%聚集、破碎并呈网格状;Vero、BHK-21细胞至第5天出现脱落、破碎和悬浮等病变,对DV有一定的敏感性;THP-1细胞至第10天未出现CPE。PCR核酸检测发现,感染DV的C6/36、THP-1、BHK和Vero细胞培养上清中可检测到DV,而Vero细胞来源DV感染的C6/36细胞未检测到DV。结论 DV对不同细胞系的感染能力和感染DV后CPE的表现形式存在较大差异。Objective To compare the CPEs of C6/36, Vero, BHK-21 and THP-1 cells caused by dengue virus（DV）and test the virus. Methods C6/36, Vero, BHK-21 and THP-1 cells were infected with DV Ⅲ strain D13053 isolated from Yunnan Province and DV Ⅱ D10156 strain from the Centers for Disease Control and Prevention of Guangdong Province, each at a MOI of 5. Meanwhile, C6/36 cells were infected with DV derived from Vero and C6/36 cells at a MOI of 5. CPE was observed until day 10 after infection, and the virus in cell culture supernatant was identified by RTPCR using the universal primers of DV. Results Of the cells, C6/36 was the most sensitive to DV, in which obvious CPE appeared, and almost all the cells were aggregated, broken or distributed as net shape. Pathological changes such as exfoliation, breakage and suspension, were observed in Vero and BHK-21 cells on day 5 after infection, and the cells showed a certain sensitivity to DV. However, no CPE was observed in THP-1 cells until day 10 after infection. DV was detected in the culture supernatants of C6/36, THP-1, BHK and Vero cells infected with DV by PCR, while undetected in C6/36 cells infected with DV derived from Vero cells. Conclusion The infective abilities of DV to various cells and the caused CPEs showed significant difference.

关 键 词：登革病毒 细胞病变 C6/36细胞 Vero细胞 BHK-21细胞 THP-1细胞 

分 类 号：R373.33[医药卫生—病原生物学] R392-33[医药卫生—基础医学]