结核分枝杆菌异烟肼或利福平不同耐药表型中sigma因子表达差异研究

Sigma factor expression in different drug-resistant phenotypes of Mycobacterium tuberculosis to isoniazid or rifampicin

  • 摘要:
    目的 分析比较在结核分枝杆菌异烟肼耐药表型和利福平耐药表型中同一种sigma因子mRNA表达量的差异。
    方法 回顾性收集天津市结核病控制中心门诊部2020—2022年耐药监测期间分离的利福平或异烟肼单耐药、耐多药和敏感菌株,提取对数期生长菌株RNA,应用荧光定量聚合酶链式反应(RT-PCR)检测结核分枝杆菌sigma因子(sigAsigL)在各个耐药组和敏感组中的mRNA表达水平,采用Mann-Whitney U检验分析各耐药组与敏感组之间的基因表达差异,采用χ2检验分析各耐药组与敏感组之间的基因高水平表达率差异。
    结果 除sigB外,sigAsigL在异烟肼单耐药组中的表达量均显著高于敏感组(均P<0.05);除sigDsigI外,sigAsigL在利福平单耐药组中的表达量均显著高于敏感组(均P<0.05);sigAsigDsigFsigGsigHsigJsigK在耐多药组(异烟肼含药培养基)与敏感组中的相对表达量比较差异有统计学意义(均P<0.05);除sigDsigI外,sigAsigL在耐多药组(利福平含药培养基)与敏感组中相对表达量比较差异有统计学意义(均P<0.05);sigGsigIsigJ、在异烟肼单耐药组中高水平表达率(高水平表达数/菌株数)均显著高于敏感组,差异有统计学意义(χ2=32.676、26.424、21.171,均P<0.05);sigJ在利福平单耐药组中高表达率高于敏感组,差异有统计学意义(χ2=21.171,P<0.05);sigGsigIsigJ在耐多药组(异烟肼含药培养基)中高表达率高于敏感组,差异有统计学意义(χ2=32.676、26.424、21.171,均P<0.05);sigJ在耐多药组(利福平含药培养基)组中高水平表达率高于敏感组,差异有统计学意义(χ2=21.171,P<0.05)。
    结论 通过对表达量和高表达率结果的综合比较和分析,sigma因子中的sigFsigGsigJ、在异烟肼单耐药、利福平单耐药、耐多药组中的表达量和表达率高于敏感组,可能与异烟肼和利福平耐药相关。

     

    Abstract:
    Objective To analyze and compare the mRNA expression levels of same sigma gene in different drug-resistant phenotypes of Mycobacterium tuberculosis.
    Methods Retrospective analysis was conducted by using the rifampicin and isoniazid resistant strains, multidrug-resistant strains, and drug sensitive strains of M. tuberculosis collected in the outpatient department of Tianjin Tuberculosis Control Center from 2018 to 2022, the RNA of M. tuberculosis was extracted in log phase, and the mRNA expression levels of M. tuberculosis sigma factor (sigA-sigL) in each drug resistant group and drug sensitive group were detected by fluorescence quantitative polymerase chain reaction (RT-PCR), and the data were analyzed by mann-whitney U test, and χ2 test was used to analyze the difference of gene high expression rate between the drug resistant groups and the drug sensitive group.
    Results Except sigB, the expression levels of sigA-sigL in the isoniazid resistant group were significantly higher than those in the drug sensitive group (P<0.05); Except sigD and sigL, the expression levels of sigA-sigL in the rifampicin resistant group were significantly higher than those in the drug sensitive group, (P<0.05). The differences in expression levels of sigA, sigD, sigF, sigG, sigH, sigJ and sigK between the multi-drug resistant group (isoniazid drug-containing medium) and the drug sensitive group were significant (all P<0.05). Except sigD and sigI, the differences in expression levels of sigA-sigL between the multi-drug resistant group (rifampicin drug-containing medium) and the drug sensitive group\ were significant (all P<0.05). The high expression rates of sigG, sigI and sigJ in isoniazid resistant groups were significantly higher than those in the drug sensitive group, the differences were significant (χ2=32.676, 26.424, 21.171, all P<0.05). The high expression rate of sigJ in rifampicin-resistant group was significantly higher than that in the drug sensitive group, the difference was significant (χ2=21.171, P<0.05). The high expression rates of sigG, sigI and sigJ in the multi-drug resistant groups (isoniazid drug-containing medium) were significantly higher than those in the drug sensitive groups, with statistical significance (χ2=32.676, 26.424, 21.171, all P<0.05). The high level expression rate of sigJ in the multi-drug resistant groups (Rifampicin drug-containing medium) was significantly higher than that in the drug sensitive group, the difference was significant (χ2=21.171, P<0.05).
    Conclusion The sigF, sigG and sigJ of M. tuberculosiswere associated with isoniazid and rifampicin resistances, indicated by their higher expression in the rifampicin resistant, isoniazid resistant and multi-drug resistant strains compared with the drug sensitive strains in this study.

     

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