The Studies And Appliance Of Single Cell Gel Electrophoresis Assay System For Detecting Mutagenicity

We applied single cell gel electrophoresis and cell culture technique, which constitute sing cell gel electrophoresis assay system for detecting mutagenicity to detect mutagenesis in vitro induced by animal drug Quinocetone and Olaquindox. And confirmed optimum lysing-time. Vero cells in the period of logarithm- growth time were treated with 9.1~273u mol/L H2O2 at 37℃ 3h, then were lysed for lh, 2h, 3h and 4h to find optimum lysing-time. Index -percentage of "comet" cell and "comettail" were analysed that indicated the optimum time is 2~3h.The degree of DNA damage by Quinocetone and Olaquindox were detected by SCGE assay, when Vero cells were treated with 1~5 ug/ml Quinocetone the percentage of live cells was above 85% and with 2-10ug/ml Olaquindox the percentage of live cells was above 90%. Vero cells in the period of logarithm- growth time were treated with 1~5ug/ml Quinocetone or 2~10u g/ml Olaquindox at 37℃ for 3h, and were lysed at 4℃ for 3h. Then were electrophored. The extent of DNA migration were measured. Index -percentage of "comet" cell and "comet tail" were analysed that indicated when Vero cells were treated with 2 u g/ml Quinocetone Vero cells got midium-grade damage, and were treated with 6 u g/ml Olaquindox Vero cells got midium-grade damage. The experimentation indicated that Quinocetone and Olaquindox can cause cellular mutation.In this paper cell culture and SCGE were applied first to detect DNA damage worked by animal drugs. And pivotal coditions is studyed.This affords technic-reservior for renewal of safety evaluation system of animal drugs.