高滴度呼肠孤病毒3型的制备及滴度检测

Preparation and titer determination of high-titer reovius 3

目的 制备高滴度呼肠孤病毒3型（reovirus 3,Reo-3）病毒液,并检测Reo-3滴度.方法 以幼仓鼠肾细胞（BHK-21）为病毒培养基质和滴度测定细胞.将Reo-3按10^0至10^-5感染复数（MOI）接种BHK-21,再分别用细胞病变法和蚀斑形成法检测病毒,用Karber法计算病毒滴度.结果 BHK-21感染Reo-3后能产生明显病变.以10^-4 MOI的Reo-3接种后48 h,收获液中病毒滴度最高,细胞病变法检测为9.625 lgTCID50/ml,蚀斑形成法检测为8.671 lgPFU/ml.结论 制备了高滴度Reo-3,为开展该病毒去除灭活研究奠定了基础.

Objective To prepare high-titer reovirus 3 （Reo-3） and determine its titer.Methods Baby hamster kidney cell （BHK-21） was used for virus cultivation and titration.The cells were infected with 10^0 to 10^-5 multiplicity of infection （MOI） of Reo-3.The virus was detected by cytopathic effect （CPE） method and plaque forming assay,respectively.Virus titers were calculated with Karber formula.Results The CPE on infected BHK-21 cells was easily observed.The highest virus titers,9.625 lgTCID50/ml tested by CPE method and 8.671 lgPFU/ml tested by plaque forming assay,were obtained when BHK-21 cells were inoculated with 10^-4 MOI of Reo-3 48 h after infection.Conclusion High titer Reo-3 can be prepared,which lays the foundation for study on virus inactivation and removal.