噬菌体TM4、Guo1和D29对静止期结核菌裂解作用初步研究

Lysis of mycobacteriophages in stationary cells of M.tuberculosis

目的研究噬菌体TM4、Guo1、D29能否裂解静止期结核菌。方法采用密闭培养至对数生长期的结核分枝杆菌构建静止期结核菌模型;采用药敏检测、金胺o-尼罗红荧光染色和电镜观察作为检测模型方法;采用most probable number(MPN)法计数作为裂解作用的检测指标。结果密闭培养11个月的静止期结核菌模型构建成功;经不同处理后,MPN计数结果为对照组1100,Guo1组23,TM4组23,D29组600,RFP组673,INH组887;与对照组相比,Guo1组和TM4组菌量明显减少(P<0.01,P<0.01),D29组菌量减少不明显(P=0.05);与RFP组相比,Guo1组和TM4组菌量亦明显减少(P<0.05,P<0.05),D29组菌量减少不明显(P>0.05);Guo1组和TM4组菌量无差异。结论噬菌体TM4和Guo1能够裂解静止期结核菌,噬菌体D29不能裂解静止期结核菌,噬菌体TM4和Guo1裂解能力无差别。

To investigate the potential of mycobacteriophages TM4, Guol and D29 in lysing stationary ceils of M. tuberculosis. Stationary cells of M. tuberculosis were induced by prolonged closed culture of logarithmic phase M. tuberculosis at 37 ℃, which were proven through drug sensitivity test, auramine o-Nile red fluorescent staining and electron microscope. The most probable number （MPN） method was used to detect whether the stationary cells of M. tuberculosis had been lysed by TM4,Guol and D29. Stationary cells of M. tuberculosis were obtained successfully by closed culture for 11m at 37 ℃. The amounts of M. tuberculosis in control, Guol, TM4,D29, RFP and INH groups by MPN method were respectively 1 100, 23, 23, 600, 673, 887 after different treatments. Compared with control group, the amount of M. tuberculosis in Guol and TM4 groups decreased more obviously（P〈0.01, P〈0.01） than 1）29 group（P=0.05）, compared with RFP group, the concentration of M. tuberculosis in Guol and TM4 groups also reduced more obviously（P〈0.05, P〈0.05） than D29 group（P〉0.05）, while there was no significant difference between the amounts of M. tuberculosis in TM4 and Guol group. In conclusion, mycobacteriophage TM4 and Guol can lyse stationary cells of M. tuberculosis, while D29 can＇t, the ability of TM4 lysing stationary cells of M. tuberculosis was the same as Guol.