一种制备复制缺陷型重组水疱性口炎病毒新方法的建立

A modified method for generation of replication-defective vesicular stomatitis virus

为了提高复制缺陷型重组水疱性口炎病毒(VSV△G*G)的制备效率,对传统制备方法进行了改良,在制备过程中先用VSV△G*G转导细胞1h后,再对其进行转染,24h后收集细胞上清并进行毒价测定。并与用传统方法制备的重组病毒的毒价进行比较,结果表明,用改良方法与传统方法制备的重组病毒具有相同的毒价,但改良方法比传统方法节省了20～48h,同时减少了表达的G蛋白诱导细胞融合所造成的细胞损伤。表明改良方法能够代替传统方法制备出高毒价的复制缺陷型重组病毒。

To improve the efficiency in generation of VSV△G·G,the traditional method was modified,BHK21 cells were transduced with VSV△G·G prior to transfection,and 1 hour later,the cells were transfected to generate new VSV△G·G.Supernatant was collected and titrated on BHK21 cells 24 hours after transfection.In result,the VSV△G·G generated by the modified method had the same titer as the recombinant virus generated by the traditional method.The modified method could save 20 to 48 hours and minimize the cell damage before harvesting VSV△G·G.In conclusion,it was feasible to conduct transduction before transfection to generate VSV△G·G.