Preparation and titer determination of high-titer reovius 3

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 100 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 104 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. Key words: Mammalian orthoreovirus 3;  Cytopathogenic effect, viral;  Plaque forming assay