论文部分内容阅读
目的建立一种快速检测胞内分枝杆菌活力的方法。方法将一定量培养至对数生长期的含pMV-eis的重组耻垢分枝杆菌感染U937巨噬细胞,以含空质粒的耻垢分枝杆菌为对照,吞噬作用2 h后洗去胞外细菌,再分别培养4、12、24和48 h后收集细胞并裂解之。获得的胞内细菌用FDA荧光染料染色后用流式细胞仪检测死亡率,并与平板菌落计数法进行比较。结果流式细胞仪检测出感染12 h后重组耻垢分枝杆菌胞内死亡率较对照组均有显著下降(P<0.05),流式细胞仪检测法与平板菌落计数法相比差异无统计学意义(P>0.05)。结论流式细胞术与传统的平板计数法相比具有快速、敏感、方便的特点,可用于分枝杆菌活菌快速检测。
Objective To establish a rapid detection of Mycobacterium intracellulare activity. Methods A certain amount of recombinant Mycobacterium smegmatis harboring pMV-eis was cultured in logarithmic growth phase to infect U937 macrophages. The plasmid containing Mycobacterium smegmatis with empty plasmid was used as a control. After phagocytosis for 2 h, the cells were washed away Bacteria, and then cultured separately 4,12,12 and 48 h after harvesting cells and lysis. The obtained intracellular bacteria were stained with FDA fluorescent dye to detect the death rate by flow cytometry and compared with the plate colony counting method. Results The intracellular cytotoxicity of recombinant M. smegmatis was significantly decreased by flow cytometry 12 h after infection (P <0.05). There was no significant difference between flow cytometry and plate colony counting Significance (P> 0.05). Conclusion Flow cytometry has the characteristics of fast, sensitive and convenient compared with the traditional plate counting method, which can be used for rapid detection of viable mycobacteria.