本文選題：結(jié)核分枝桿菌　切入點(diǎn)：耐藥性　出處：《石河子大學(xué)》2014年碩士論文　論文類型：學(xué)位論文

【摘要】：目的： 通過比較分析不同耐藥性的結(jié)核分枝桿菌臨床分離株P(guān)hoP基因和PhoR基因表達(dá)水平的差異，，探討研究結(jié)核分枝桿菌PhoPR雙組分系統(tǒng)與新疆地區(qū)廣泛流行的耐藥結(jié)核桿菌耐藥性是否具有相關(guān)性。 方法： 以新疆地區(qū)廣泛流行的耐藥結(jié)核分枝桿菌臨床分離株為研究對象，分別提取結(jié)核分枝桿菌臨床分離藥物敏感菌株、單純耐異煙肼的結(jié)核分枝桿菌臨床分離株(INH-MTB)、單純耐利福平的結(jié)核分枝桿菌臨床分離株（RFP-MTB）、單純耐鏈霉素的結(jié)核分枝桿菌臨床分離株(SM-MTB)、單純耐乙胺丁醇的結(jié)核分枝桿菌臨床分離株(EB-MTB)、耐多藥的結(jié)核分枝桿菌臨床分離株的結(jié)核分枝桿菌菌株(MDR)的總RNA，并進(jìn)行純度鑒定。運(yùn)用SYBR Green I實(shí)時(shí)熒光定量PCR的方法，分別檢測各組結(jié)核分枝桿菌菌株原始狀態(tài)下的PhoP基因和PhoR基因的表達(dá)，比較分析不同耐藥性的結(jié)核分枝桿菌臨床分離株P(guān)hoP基因和PhoR基因表達(dá)水平的差異。檢測比較分析各組結(jié)核分枝桿菌在INH、RFP、SM、EB抗結(jié)核藥物選擇壓力下，各組結(jié)核分枝桿菌菌株的PhoP基因和PhoR基因表達(dá)的變化；檢測比較分析各組結(jié)核分枝桿菌在復(fù)制的感染小鼠動(dòng)物模型的肺臟、肝臟和脾臟內(nèi)，各組結(jié)核分枝桿菌菌株的PhoP基因和PhoR基因表達(dá)的變化。 結(jié)果： 1.在原始狀態(tài)下，耐藥性結(jié)核分枝桿菌臨床分離株的PhoP基因和PhoR基因的mRNA表達(dá)水平比臨床分離藥物敏感菌株顯著升高，差異具有統(tǒng)計(jì)學(xué)意義（P0.05）； 2.在抗結(jié)核藥物（INH、RFP、SM、EB）選擇壓力作用下，結(jié)核分枝桿菌PhoP基因和PhoR基因的mRNA表達(dá)水平均較各組結(jié)核分枝桿菌在原始狀態(tài)下表達(dá)上調(diào)，差異具有統(tǒng)計(jì)學(xué)意義（P0.05）； 3.在各組結(jié)核分枝桿菌感染小鼠動(dòng)物模型的肺臟、肝臟和脾臟內(nèi)，各組耐藥結(jié)核分枝桿菌臨床分離株的PhoP基因和PhoR基因的mRNA表達(dá)水平比各組結(jié)核分枝桿菌在原始狀態(tài)下顯著升高，差異具有統(tǒng)計(jì)學(xué)意義（P0.05）。 結(jié)論： 1.在結(jié)核分枝桿菌臨床分離藥物敏感菌株、INH-MTB、RFP-MTB、SM-MTB、EB-MTB和MDR中，結(jié)核分枝桿菌PhoP和PhoR基因的表達(dá)有差異； 2.在抗結(jié)核藥物（INH、RFP、SM、EB）選擇壓力作用下，各組耐藥結(jié)核分枝桿菌臨床分離株的PhoP基因和PhoR基因的mRNA表達(dá)水平比各組結(jié)核分枝桿菌在原始狀態(tài)下顯著升高； 3.在各組結(jié)核分枝桿菌感染小鼠動(dòng)物模型的肺臟、肝臟和脾臟內(nèi)，各組耐藥結(jié)核分枝桿菌臨床分離株的PhoP基因和PhoR基因的mRNA表達(dá)水平比各組結(jié)核分枝桿菌在原始狀態(tài)下顯著升高；綜上所述：結(jié)核分枝桿菌PhoPR雙組分系統(tǒng)與新疆地區(qū)廣泛流行的耐藥結(jié)核分枝桿菌的耐藥性具有相關(guān)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.
【學(xué)位授予單位】：石河子大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2014
【分類號】：R378.911

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