Establishment of determination of murine minute virus and preparation of the virus by response surface methodology
[Objective]To develop a method for determination of infectious titer of murine minute virus(MMV)and prepare high titer MMV.[Method]The Box-Behnken based optimization was used to prepare MMV with A9 cell and a median infec-tive dose of cell culture(TCID50)method for determination of MMV titer.[Result]The optimal A9 cell concentration for inocu-lation,virus culture time and virus adsorption time were 9 × 103cells/well,12 d and 0 h respectively.The developed TCID50 method showed good precision.The MMV titer of A9 cells in precipitate was significantly higher than that in supernatant,and increased gradually with the increasing MOI,which reached the peak value 72 h after infection with a MOI of 0.5.[Conclu-sion]The Box-Behnken could use in MMV with A9 cell and a median infective dose of cell culture(TCID50)method for deter-mination of MMV titer.The mean tite of prepared MMV was 6.35 TCID50/0.1 mL.
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