流式细胞术结合荧光标记技术检测洗必泰对体外粪肠球菌生物膜的作用  

Application of flow cytometry combined with fluorescent labeling technique to detect the effect of chlorhexidine on E. faecalis biofilm in vitro

作　　者：丁晓晓 杜田丰[2] 唐学智[1] 姚怡辰 张千 宋珂[1] 曹颖光[1] DING Xiao-xiao;DU Tian-feng;TANG Xue-zhi;YAO Yi-chen;ZHANG Qian;SONG Ke;CAO Ying-guang(Faculty and Department of Stomatology, Tongji Hospital, Huazhong University of Science and Technology,Hubei Wuhan 430030, China;Department of Stomatology The First Affiliated Hospital of Zhengzhou University,Henan Zhengzhou 450052, China)

机构地区：[1]华中科技大学同济医学院附属同济医院口腔医学中心,湖北武汉430030 [2]郑州大学第一附属医院口腔医学中心,河南郑州450052

出　　处：《临床口腔医学杂志》2019年第5期267-271,共5页Journal of Clinical Stomatology

基　　金：国家自然科学基金项目(81500847)

摘　　要：目的:利用流式细胞术结合荧光标记技术检测洗必泰对体外粪肠球菌(Enterococcus faecalis,E.faecalis生物膜的作用。方法:应用2%洗必泰(CHX)及生理盐水溶液(NS)处理体外已培养1 d和3 w的粪肠球菌生物膜样本5 min,随后在培养的第0 d、1 d和7 d经PI和SYTO 9共同染色后运用流式细胞仪检测观察2%CHX的灭菌效果。结果:2%CHX针对体外粪肠球菌生物膜的灭菌效果优于生理盐水组且差异有统计学意义(P<0. 05),且2%CHX对培养3 w生物膜的杀灭效果较对培养1 d生物膜的杀灭效果减弱,差异有统计学意义(P<0. 05)。培养1 d、3w的生物膜经2%CHX处理后随着再培养时间的延长,细菌中活菌比例逐渐增多,且再培养1 w后活菌所占比例与生理盐水组之间的差异无统计学意义(P>0. 05)。结论:2%洗必泰溶液对体外不同生长阶段的粪肠球菌生物膜均具有良好的灭菌效果,但灭菌效果不彻底。Objective: To detect the effect of chlorhexidine on the biofilm of Enterococcus faecalis( E.faecalis) in vitro by flow cytometry( FCM). Methods: Both 1 d and 3 w E. faecalis biofilms were cultured in vitro,and biofilm samples were treated with 2% CHX and normal saline solution( NS) for 5 min,then,after culturing for 0 d,1 d and 7 d,the sterilizing effect was observed by flow cytometry after co-staining with PI and SYTO 9.Results: The sterilizing effect against E.faecalis biofilm of 2% CHX was stronger than the saline group with statistically significance( P<0. 05).The killing effect of 2% CHX on cultured 3 w biofilm weakened compared with the killing effect on cultured 1 d biofilm significantly( P< 0. 05).The longer biofilms cultured after treated with 2% CHX,the more proportion of live bacteria in the bacteria appeared.Besides the difference of the proportion of live bacteria between saline group and after 1 w culture was not statistically significant( P>0. 05).Conclusion: The 2% chlorhexidine solution has a good sterilizing effect on the E. faecalis biofilm in different growth stages in vitro,and the sterilizing effect is not completely.

关 键 词：洗必泰溶液 粪肠球菌生物膜 流式细胞术 灭菌 

分 类 号：R781.05[医药卫生—口腔医学]