qRT-PCR检测结核分枝杆菌毒素-抗毒素系统mazE F的表达

Detection on expression levels of mazE F toxin-antitoxin system in Mycobacterium tuberculosis by qRT-PCR

目的探讨结核分枝杆菌毒素基因mazF3,6,9及抗毒素基因mazE3,6,9的表达差异。方法运用实时定量PCR检测结核分枝杆菌单耐药株20株,耐多药株20株和标准株H37Rv毒素基因mazF3,6,9及抗毒素基因mazE3,6,9的表达水平;组间基因表达水平的差异用one-way ANOVA进行统计分析。结果与对照株相比较,毒素基因mazF6,9在单耐药组(11.151 9±22.317 21;8.430 6±17.978 97)及耐多药组(4.601 6±1.290 18;6.962 7±6.929 48)表达均上调且差异有统计学意义(P<0.01),mazF3基因在单耐药组及耐多药组差异无统计学意义(P>0.05),抗毒素基因mazE3在单耐药(0.360 6±0.125 27)及耐多药组(0.201 6±0.165 42)中表达均下调,差异有统计学意义(P<0.01),mazE6均无统计学意义,mazE9只有在耐多药组(0.398 9±0.376 79)中表达下调,差异有统计学意义,(P<0.01)。结论毒素基因mazF6,9抗毒素基因mazE3,9可能参与了结核分枝杆菌的耐药形成,具体机制尚待进一步的研究。

We investigate the different expression of toxin gene mazF3,6,9 and antitoxin gene mazE3,6,9 in the drug-re- sistance Mycobacterium tuberculosis, we used quantitative real-time polymerase chin reaction method to detect the expression level of toxin gene rnazF3,6,9 and antitoxin gene mazE3,6,9 in M. tuberculosis （20 mono-resistance strains, 20 muhidrug re- sistance strains and standard strain H37Rv）. The differences of gene expression levels between groups were analyzed by one- way ANOVA. Contrasting with control group, toxin genes rnazF6,9 were up-regulated expression levels both in mono-resistance （11. 1519±22. 31721 ;8. 4306±17. 97897） and multidrug resistance （4. 6016±1. 29018;6. 9627±6. 92948）, had statisti- cal significance （P〈0.01）, mazF3 expression levels had statistical significance neither in mono-resistance nor in rnultidrug re- sistance （P〉0.05） ; antitoxin genes mazE3 was in down-expression level, and had statistical significance both in mono-resist- ance （0. 3606±0. 12527） and multidrug resistance （0. 2016±0. 16542） （P〈0.01）, mazE6 had no statistical significance （P〉 0.05）either in mono-resistance or multi drug resistance, mazE9 only in multidrug resistance（0. 3989±0. 37679） was in down-expression level, and has statistical significance （P〈0. 001）. The toxin gene mazF6,9 and antitoxin gene mazE3,9 may participate in the drug-resistance formation of M. tuberculosis.