流式细胞术快速检测白色念珠菌对伊曲康唑敏感性方法的研究

Study on mothed of detecting susceptibility of candida albicans to itraconazole by rapid flow cytometry

目的:建立流式细胞术快速检测白色念珠菌对伊曲康唑敏感性试验(FCST)的方法。方法:以碘化丙锭(PI)为荧光染料,使用活菌和乙醇处理的死菌在SS/log(FL3)图上进行设门,用流式细胞术检测标准菌株的死菌比率。使用标准菌株对本法与CLSI常量稀释法进行平行测定,得出本法的M IC判断标准。同时用此法对30株临床菌株进行双份平行检测并与NCCLS常量稀释法(M27-A)进行比较。结果:经过调整电压把活菌与死菌在SS/log(FL3)图上分为两群分界清晰的细胞。对30株临床分离菌株的检测发现,本法具有很好的重复性,与M27-A比较,结果无显著性差异(P=0.141)。结论:本法具有快速、敏感、可靠的优势,在临床抗真菌药物敏感性检测中具有很大的应用前景。

Objective：To establish a method to detect susceptibility of Candida albicans to itraconazole by the flow cytometry.Methods：Dyed by PI fluorescent dye and gated with viable bacteria and killed bacteria prepared with ethanol on the figure of SS/log（FL3）,the flow cytometry was used to detect the rate of killed bacterium in standard bacterial stain.The standard value of MIC was acquired according to the parallel determination between the flow cytometry and CLSI broth macrodilution method in standard bacterial stain.Simultaneously,the flow cytometry was used to detect 30 clinical stains in double and parallel way and be compared with the NCCLS broth microdilution method（M27-A）.Results：The viable bacteria and killed bacteria were clearly divided into two groups on the figure of SS/log（FL3） by adjusting voltage.After detecting 30 clinical bacterium stains,we discovered that the flow cytometry was very repeatable.It was found that no significant different existed between the flow cytometry and M27-A（P=0.141）.Conclusion：The method of FCST we established in this study is simple,repeatable and of high accuracy and likely to determine MIC and has vast application prospects in clinical antifungal susceptibility testing.