猪圆环病毒2型基因1群B亚簇的体外培养特征  

In vitro replication characteristics of porcine circovirus 2(PCV2)belonging to B subcluster of genogroup1

作　　者：罗维[1] 赵墩[1] 吴海超[1] 蒋大良[1] 余兴龙[1] 

机构地区：[1]湖南农业大学动物医学院,湖南长沙410128

出　　处：《湖南农业大学学报（自然科学版）》2013年第5期534-538,共5页Journal of Hunan Agricultural University(Natural Sciences)

基　　金：国家自然科学基金项目(30571390)

摘　　要：为探寻猪圆环病毒2型(PCV2)在PK15细胞中的复制特征,将PCV2基因1群B亚簇(PCV2–1B)病毒感染PK15均质细胞A10,待感染细胞层长满,对其进行连续传代;对第5、6代细胞进行维持培养,即在细胞长满后隔天更换培养基,以维持细胞良好的生长;用第10代带毒细胞的培养液接种A10细胞,并进行换液维持培养。PCR与定量PCR检测结果表明,PCV2–1B病毒感染A10细胞后,其病毒基因组的含量在感染细胞长满之前(约96 h)随着细胞的增多而上升,同时也随着传代次数的增多而上升;换液维持培养的细胞,在维持10 d后PCV2–1B病毒基因组的含量能维持在107～108拷贝/mL,而第10代带毒细胞的培养液接毒A10之后,病毒基因组含量随细胞增多而上升,在达到约108拷贝/mL后,病毒基因组含量维持在107～108拷贝/mL。To observe the replication dynamics of PCV2 in PK15 cell, PCV2-1B infectious clone virus was inoculated to the homogenous PK15（A10） cells. After grew to 100% confluence, the cell was serially passaged. And part of the 5th and 6th -generation cells were maintained by altering cell culture medium at one day interval to maintain the cells. The culture medium of the 10th generation cells was inoculated to fresh A10 cells which was also maintained by culture medium alteration. The results of PCR and quantitative PCR showed that after infection, the PCV2 genome copies increased with the growth of the cells until the cell reached 100% confluence. And the PCV2 genome copies maintained at a level of 107-108 copies/mL in cells after maintained by culture medium alteration for 10 days. Infection of A10 cell with culture medium of the 10th generation cells also showed the virus genome content increased with the growth of the cell, and when reached 108 copies/mL, the genome copies maintained at 107-108 copies/mL.

关 键 词：猪圆环病毒2型 复制动态 PK15细胞 毒株 

分 类 号：S852.659[农业科学—基础兽医学]