结核杆菌PhoPR双组分系统与新疆地区耐药结核杆菌临床分离株耐药性的相关性研究

发布时间：2018-03-23 09:05

  本文选题：结核分枝杆菌　切入点：耐药性　出处：《石河子大学》2014年硕士论文　论文类型：学位论文

【摘要】：目的： 通过比较分析不同耐药性的结核分枝杆菌临床分离株PhoP基因和PhoR基因表达水平的差异，，探讨研究结核分枝杆菌PhoPR双组分系统与新疆地区广泛流行的耐药结核杆菌耐药性是否具有相关性。 方法： 以新疆地区广泛流行的耐药结核分枝杆菌临床分离株为研究对象，分别提取结核分枝杆菌临床分离药物敏感菌株、单纯耐异烟肼的结核分枝杆菌临床分离株(INH-MTB)、单纯耐利福平的结核分枝杆菌临床分离株（RFP-MTB）、单纯耐链霉素的结核分枝杆菌临床分离株(SM-MTB)、单纯耐乙胺丁醇的结核分枝杆菌临床分离株(EB-MTB)、耐多药的结核分枝杆菌临床分离株的结核分枝杆菌菌株(MDR)的总RNA，并进行纯度鉴定。运用SYBR Green I实时荧光定量PCR的方法，分别检测各组结核分枝杆菌菌株原始状态下的PhoP基因和PhoR基因的表达，比较分析不同耐药性的结核分枝杆菌临床分离株PhoP基因和PhoR基因表达水平的差异。检测比较分析各组结核分枝杆菌在INH、RFP、SM、EB抗结核药物选择压力下，各组结核分枝杆菌菌株的PhoP基因和PhoR基因表达的变化；检测比较分析各组结核分枝杆菌在复制的感染小鼠动物模型的肺脏、肝脏和脾脏内，各组结核分枝杆菌菌株的PhoP基因和PhoR基因表达的变化。 结果： 1.在原始状态下，耐药性结核分枝杆菌临床分离株的PhoP基因和PhoR基因的mRNA表达水平比临床分离药物敏感菌株显著升高，差异具有统计学意义（P0.05）； 2.在抗结核药物（INH、RFP、SM、EB）选择压力作用下，结核分枝杆菌PhoP基因和PhoR基因的mRNA表达水平均较各组结核分枝杆菌在原始状态下表达上调，差异具有统计学意义（P0.05）； 3.在各组结核分枝杆菌感染小鼠动物模型的肺脏、肝脏和脾脏内，各组耐药结核分枝杆菌临床分离株的PhoP基因和PhoR基因的mRNA表达水平比各组结核分枝杆菌在原始状态下显著升高，差异具有统计学意义（P0.05）。 结论： 1.在结核分枝杆菌临床分离药物敏感菌株、INH-MTB、RFP-MTB、SM-MTB、EB-MTB和MDR中，结核分枝杆菌PhoP和PhoR基因的表达有差异； 2.在抗结核药物（INH、RFP、SM、EB）选择压力作用下，各组耐药结核分枝杆菌临床分离株的PhoP基因和PhoR基因的mRNA表达水平比各组结核分枝杆菌在原始状态下显著升高； 3.在各组结核分枝杆菌感染小鼠动物模型的肺脏、肝脏和脾脏内，各组耐药结核分枝杆菌临床分离株的PhoP基因和PhoR基因的mRNA表达水平比各组结核分枝杆菌在原始状态下显著升高；综上所述：结核分枝杆菌PhoPR双组分系统与新疆地区广泛流行的耐药结核分枝杆菌的耐药性具有相关性。
[Abstract]:Objective:. The expression levels of PhoP gene and PhoR gene in clinical isolates of Mycobacterium tuberculosis with different drug resistance were compared and analyzed. To investigate the correlation between the PhoPR two-component system of Mycobacterium tuberculosis and the drug resistance of drug-resistant Mycobacterium tuberculosis in Xinjiang. Methods:. The clinical drug sensitive strains of Mycobacterium tuberculosis were isolated from the clinical isolates of Mycobacterium tuberculosis, which are widely prevalent in Xinjiang. Isoniazid resistant Mycobacterium tuberculosis clinical isolates INH-MTBN, rifampicin-resistant Mycobacterium tuberculosis clinical isolates RFP-MTBN, Streptomycin-resistant Mycobacterium tuberculosis clinical isolates SM-MTBN, simple ethambutanol-resistant tuberculous branching rods. The total RNAs and purity of EB-MTBN, a multidrug resistant mycobacterium tuberculosis clinical isolates, were identified by SYBR Green I real-time fluorescence quantitative PCR. The expression of PhoP gene and PhoR gene in the original condition of Mycobacterium tuberculosis strains were detected. The expression levels of PhoP gene and PhoR gene in clinical isolates of Mycobacterium tuberculosis with different drug resistance were compared and analyzed. The changes of PhoP gene and PhoR gene expression of Mycobacterium tuberculosis strains in each group were compared and analyzed in the lungs, liver and spleen of the infected mice. Changes of PhoP gene and PhoR gene expression of Mycobacterium tuberculosis strains in each group. Results:. 1. In the original condition, the mRNA expression level of PhoP gene and PhoR gene in clinical isolates of drug-resistant Mycobacterium tuberculosis was significantly higher than that of drug sensitive strains, and the difference was statistically significant (P 0.05). 2. The mRNA expression level of PhoP gene and PhoR gene of Mycobacterium tuberculosis was significantly higher than that of Mycobacterium tuberculosis in the original condition under the selective pressure of anti-tuberculosis drug INHS-RFPS-SM-EB.The difference was statistically significant (P 0.05). 3. In the lung, liver and spleen of mice infected with Mycobacterium tuberculosis, The mRNA expression level of PhoP gene and PhoR gene in the clinical isolates of Mycobacterium tuberculosis was significantly higher than that in the original state of Mycobacterium tuberculosis, and the difference was statistically significant (P 0.05). Conclusion:. 1. The expression of PhoP gene and PhoR gene in Mycobacterium tuberculosis were different in the clinical isolates of Mycobacterium tuberculosis, INH-MTBU RFP-MTB, SM-MTBU EB-MTB and MDR. 2. The mRNA expression level of PhoP gene and PhoR gene in clinical isolates of drug-resistant Mycobacterium tuberculosis was significantly higher than that of Mycobacterium tuberculosis in the original condition. 3. In the lung, liver and spleen of mice infected with Mycobacterium tuberculosis, The mRNA expression levels of PhoP gene and PhoR gene in the clinical isolates of Mycobacterium tuberculosis were significantly higher than those in each group. To sum up, the PhoPR two-component system of Mycobacterium tuberculosis is related to the drug resistance of Mycobacterium tuberculosis, which is widely prevalent in Xinjiang.
【学位授予单位】：石河子大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R378.911

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