【作者】 高鸣； 谭耀驹； 谭学端； 曹翌明； 关平；

【Author】 Gao ming,Tan yaoju,Tan xueduan.Cao yimin,Guan ping Department of Clinical Laboratory,,Guangzhou Chest Hospital,Guangzhou,510095,China

【机构】 广州市胸科医院检验科；

【摘要】 目的探讨建立利用流式细胞仪检测结核分枝杆菌在肺泡上皮A549细胞内药敏实验的方法。方法利用标记了乙酰乙酸荧光素FDA的结核分枝杆菌感染肺泡上皮A549细胞,构建成侵染模型。用阿米卡星杀死细胞外的结核分枝杆菌。再用左氧氟沙星与感染结核分枝杆菌的细胞孵育,以作用于细胞内的结核分枝杆菌。利用流式细胞仪检测氧氟沙星作用前后细胞内结核分枝杆菌的荧光强度的变化,从而检测出结核分枝杆菌在细胞内的药敏情况。结果感染细胞的20株结核分枝杆菌在左氧氟沙星作用前后荧光强度无明显变化(p>0.05)。结论目前流式细胞仪虽可以快速检测结核分枝杆菌的药敏情况,但是对于细胞内结核分枝杆菌药敏检测由于存在染料残留产生荧光干扰等问题,还未能找到较好的方法检测结核分枝杆菌在细胞内的药敏情况。更多还原

【Abstract】 Objective:To discuss the feasibility of detecting drug sensitivity of mycobacterium tuberculosis in A549 cell using flow cytometry. Methods The method was based on the ability of viable mycobacterium organisms to hydrolyze fluorescein diacetate(FDA).First,A549 cells were infected with mycobacterium tuberculosis labeled FDA.Second,the extracellular mycobacterium tuberculosis was killed by amikacin,then the infected cells were incubated with levofloxacin.Comparing the intensity of fluorescence of mycobacterium cells incubated with levofloxacin with that of levofloxacin-free cells. Results The intensity of fluorescence of levofloxacin cells did not change significantly(p>0.05).There was no statistical significance. Conclusion The use of flow cytometry has already show considerable promise as a rapid method for obtaining susceptibility test tesults.However,because of some existed problem such as dye residual,there was no better method to analyse the drug susceptibility of intracellular Mycobacterium tuberculosis.更多还原