發(fā)布時間：2018-05-13 04:10

  本文選題：MIRU + 結(jié)核��； 參考：《皖南醫(yī)學(xué)院》2016年碩士論文

【摘要】：目的：MIRU位點是否同結(jié)核耐藥存在相關(guān)性,以及同哪些抗結(jié)核藥物存在相關(guān)性的研究相對較少。本次研究的目的在于評價MTUB21等21個MIRU位點與異煙肼(INH)、利福平(RFP)、鏈霉素(SM)、乙胺丁醇(EMB)及吡嗪酰胺(PSA)耐藥的相關(guān)性。方法：將搜集的結(jié)核菌株進(jìn)行痰涂片檢查,選痰涂片陽性樣本分別接種于酸性改良羅氏培養(yǎng)基、對硝基苯甲酸鑒別培養(yǎng)基、噻吩-2-羧酸肼培養(yǎng)基。選擇三個培養(yǎng)基均陽性的樣本(即結(jié)核分枝桿菌)進(jìn)行INH、RFP、SM、EMB及PSA的藥敏實驗。將上述樣本提取總DNA作為模板,通過PCR擴(kuò)增擴(kuò)增出MTUB21等15個MIRU位點的序列并進(jìn)行凝膠電泳。利用凝膠電泳條帶并通過MIRU位點拷貝次數(shù)讀數(shù)表確定每個樣本的MIRU位點的拷貝次數(shù)。統(tǒng)計分析通過SPSS16.0完成。利用描述性統(tǒng)計方法描述105個結(jié)核分枝桿菌菌株的耐藥情況及不同拷貝次數(shù)的耐藥頻率分布。單因素和多因素logistic回歸分析來評價15個MIRU位點和結(jié)核耐藥的相關(guān)性。此外,不同MIRU位點的遺傳性差異和分辨率通過Mircosoft Excel完成。P0.05視為具有統(tǒng)計學(xué)意義。結(jié)果：15個MIRU位點的遺傳性差異從大到小依次是MTUB21MTUB04 MIRU26QUB11bMIRU 16MIRU39MIRU20MIRU27ETREMIRU10MIRU4 0ETRAETRDETRBETRC;經(jīng)過多因素logistic回歸分析結(jié)果發(fā)現(xiàn),INH耐藥的抑制性位點為ETRB(P=0.022, OR=0.104,95%CI:0.015-0.718); RFP耐藥的抑制性位點為MTUB21(P=0.018, OR=0.560,95%CI:0.347-0.906); SM的促進(jìn)性位點為MIRU20(P=0.049, OR=9.162,95%CI:1.011-83.015),其抑制性位點為MTUB21(P=0.024,OR=0.425,95% CI:0.203-0.891)；EMB的抑制性位點為MIRU 27 (P=0.040,OR=0.299,95%CI:0.095-0.945)；PSA的抑制性位點為ETRA (P=0.02 4,OR=0.396,95%CI:0.177-0.887).結(jié)論：不同MIRU位點的分辨率存在差異,部分MIRU位點與結(jié)核耐藥存在相關(guān)性(ETRB和INH耐藥；MIRU21、MTUB21和RFP耐藥；MIRU27和EMB耐藥；ETRA和PSA耐藥)�？赡艽嬖诘臋C(jī)制為MIRU位點上調(diào)或下調(diào)了位點附近功能基因的轉(zhuǎn)錄,引起分枝菌酸等多種成分表達(dá)的減少或增加,并進(jìn)一步對結(jié)核分枝桿菌是否耐藥造成影響。
[Abstract]:Objective there is relatively little research on whether the 1: MIRU locus is associated with TB resistance and with which anti-TB drugs. The aim of this study was to evaluate the association of 21 MIRU sites including MTUB21 with isoniazid, rifampicin, streptomycin, ethambutanol and pyrazinamide. Methods: the collected strains of tuberculous bacilli were examined for sputum smear. The positive samples were inoculated in acid modified Roche medium, p-nitrobenzoic acid identification medium and thiophene -2-carboxylic acid hydrazine medium respectively. Three positive samples (Mycobacterium tuberculosis) were selected to test the drug sensitivity of PSA and SMMB. The total DNA was extracted from the above samples as template, and 15 MIRU sites such as MTUB21 were amplified by PCR and gel electrophoresis was carried out. The copy times of MIRU loci in each sample were determined by the MIRU site copy number reading table using the gel electrophoresis strip. The statistical analysis was completed by SPSS16.0. The drug resistance of 105 strains of Mycobacterium tuberculosis and the frequency distribution of drug resistance in different copies were described by descriptive statistical method. Univariate and multivariate logistic regression analysis was used to evaluate the correlation between 15 MIRU loci and TB resistance. In addition, the genetic differences and resolution of different MIRU loci were statistically significant through Mircosoft Excel completion. Results: the genetic difference of 15 MIRU loci from large to small was MTUB21MTUB04 MIRU26QUB11bMIRU 16MIRU39MIRU20MIRU27ETREMIRU10MIRU4 0ETRAETRDETRBETRC.The results of multivariate logistic regression analysis showed that the inhibitory sites of MIRU resistance were ETRBP 0.022, ORX 0.10495CI: 0.015-0.718; RFP resistance was MTUB21PU 0.018, ORUB0.56095 CIW 0.347-0.906; SM's promotive locus was 0.347-0.906. The inhibitory sites of MIRU20 / P0.049, OR9.162 / 95CI1: 1.011-83.015, and the inhibitory sites of MTUB21P0. 02595% CI0. 203-0. 891EMB are MIRU 27, P0.040, OR0. 29995, CI0.095-0.945. The inhibitory sites of ETRA P0.02 4OR0. 39695 are 0.177-0.887. Conclusion: there are differences in the resolution of different MIRU loci, and some MIRU loci are correlated with TB resistance. The drug resistance of INH and MIRU21 MTUB21 and RFP resistance is MIRU27 and EMB resistance. The possible mechanism is that MIRU upregulated or down-regulated the transcription of functional genes near the locus, resulting in the decrease or increase of the expression of mycoic acid and other components, and further affected the drug resistance of Mycobacterium tuberculosis.
【學(xué)位授予單位】：皖南醫(yī)學(xué)院
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2016
【分類號】：R52

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1 蔣超;MTUB21等MIRU位點與結(jié)核耐藥的相關(guān)性研究[D];皖南醫(yī)學(xué)院;2016年

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本文編號：1881592