3种中和剂对常用消毒剂在Vero细胞上的中和效果

Neutralizing effects of 3 neutralizers on common disinfectants in Vero cells

目的 研究0.5%硫代硫酸钠、10%胰蛋白胨大豆肉汤(tryptic soy broth,TSB)培养基、10% Letheen肉汤培养基对实验室常用的4种消毒剂的中和效果,为疫苗生产和检定中科学选择中和剂提供依据.方法 取生长良好的Vero细胞接种于96孔培养板,每组8个复孔,置于37℃、5% CO2培养箱培养过夜.将3种中和剂分别与杀孢子剂、0.5%“84”消毒液、酸性苯酚盐、碱性苯酚盐4种消毒剂中和后,接种于单层生长的Vero细胞,显微镜下观察细胞生长情况.结果 0.5%硫代硫酸钠分别与杀孢子剂、0.5%“84”消毒液中和后接种细胞,8孔细胞全部存活;而0.5%硫代硫酸钠分别与酸性苯酚盐和碱性苯酚盐的中和产物接种细胞后,细胞全部死亡.10% TSB培养基分别与酸性苯酚盐、碱性苯酚盐中和后接种细胞,8孔细胞全部存活;而10% TSB培养基分别与杀孢子剂和0.5%“84”消毒液的中和产物接种细胞后,细胞全部死亡.10% Letheen肉汤培养基接种细胞后,8个孔细胞全部死亡.结论 0.5%硫代硫酸钠可中和杀孢子剂和0.5%“84”消毒液的消毒作用,10% TSB培养基可中和酸性苯酚盐和碱性苯酚盐的消毒作用.10% Letheen肉汤培养对Vero细胞有毒性作用.

Objective To investigate the neutralizing effects of 0.5% sodium thiosulfate,10% tryptic soy broth (TSB) medium,10% Letheen broth medium on 4 disinfectants commonly used in laboratories,and provide a basis for neutralizer selection in vaccine production and verification.Methods Well-grown Vero cells were seeded in 96-well plates,with 8 replicate wells per group,and cultured overnight at 37 ℃,5% CO2.The above 3 neutralizers were mixed with 4 disinfectants-sporicide,0.5% "84" disinfectant,acidic phenolate and basic phenolate,respectively.Then the neutralized products were inoculated with single-layered Vero cells.The cell morphology was observed under microscope.Results When the products of sporicide or 0.5% "84" disinfectant neutralized by 0.5% sodium thiosulfate were inoculated with Vero cells,all 8-well cells survived,yet when the products of acidic phenolate or basic phenolate neutralized by 0.5% sodium thiosulfate were inoculated with the cells,all cells died.When the products of acidic phenolate or basic phenolate neutralized by 10% TSB medium were inoculated with the cells,all 8-well cells survived,yet when the products of sporicide or 0.5% "84" disinfectant neutralized by 10% TSB medium were inoculated with the cells,all cells died.After 10% Letheen broth medium was inoculated with Vero cells,all 8-well cells died.Conclusions 0.5 % sodium thiosulfate can neutralize sporicide and 0.5% “84” disinfectant,and 10% TSB medium can neutralize acidic phenolate and basic phenolate.10% Letheen broth medium has toxic effect on Vero cells.