3种中和剂对常用消毒剂在Vero细胞上的中和效果

 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培养过夜。将3种中和剂分别与杀孢子剂、0.5%“84”消毒液、酸性苯酸盐、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。

Neutralizing effect of 3 neutralizers on common disinfectants in Vero cells

Objective  To investigate the neutralizing effect 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 into 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 into 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 into the cells, all cells died. When the products of acidic phenolate and basic phenolate neutralized by 10% TSB medium were inoculated into the cells, all 8-well cells survived, yet when the products of sporicide or 0.5% "84" disinfectant separately neutralized by 10% TSB medium were inoculated into the cells, all cells died. After 10% Letheen broth medium was inoculated into 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.