MTUB21等MIRU位点与结核耐药的相关性研究

发布时间：2018-05-13 04:10

本文选题：MIRU + 结核　；参考：《皖南医学院》2016年硕士论文

【摘要】：目的：MIRU位点是否同结核耐药存在相关性,以及同哪些抗结核药物存在相关性的研究相对较少。本次研究的目的在于评价MTUB21等21个MIRU位点与异烟肼(INH)、利福平(RFP)、链霉素(SM)、乙胺丁醇(EMB)及吡嗪酰胺(PSA)耐药的相关性。方法：将搜集的结核菌株进行痰涂片检查,选痰涂片阳性样本分别接种于酸性改良罗氏培养基、对硝基苯甲酸鉴别培养基、噻吩-2-羧酸肼培养基。选择三个培养基均阳性的样本(即结核分枝杆菌)进行INH、RFP、SM、EMB及PSA的药敏实验。将上述样本提取总DNA作为模板,通过PCR扩增扩增出MTUB21等15个MIRU位点的序列并进行凝胶电泳。利用凝胶电泳条带并通过MIRU位点拷贝次数读数表确定每个样本的MIRU位点的拷贝次数。统计分析通过SPSS16.0完成。利用描述性统计方法描述105个结核分枝杆菌菌株的耐药情况及不同拷贝次数的耐药频率分布。单因素和多因素logistic回归分析来评价15个MIRU位点和结核耐药的相关性。此外,不同MIRU位点的遗传性差异和分辨率通过Mircosoft Excel完成。P0.05视为具有统计学意义。结果：15个MIRU位点的遗传性差异从大到小依次是MTUB21MTUB04 MIRU26QUB11bMIRU 16MIRU39MIRU20MIRU27ETREMIRU10MIRU4 0ETRAETRDETRBETRC;经过多因素logistic回归分析结果发现,INH耐药的抑制性位点为ETRB(P=0.022, OR=0.104,95%CI:0.015-0.718); RFP耐药的抑制性位点为MTUB21(P=0.018, OR=0.560,95%CI:0.347-0.906); SM的促进性位点为MIRU20(P=0.049, OR=9.162,95%CI:1.011-83.015),其抑制性位点为MTUB21(P=0.024,OR=0.425,95% CI:0.203-0.891)；EMB的抑制性位点为MIRU 27 (P=0.040,OR=0.299,95%CI:0.095-0.945)；PSA的抑制性位点为ETRA (P=0.02 4,OR=0.396,95%CI:0.177-0.887).结论：不同MIRU位点的分辨率存在差异,部分MIRU位点与结核耐药存在相关性(ETRB和INH耐药；MIRU21、MTUB21和RFP耐药；MIRU27和EMB耐药；ETRA和PSA耐药)。可能存在的机制为MIRU位点上调或下调了位点附近功能基因的转录,引起分枝菌酸等多种成分表达的减少或增加,并进一步对结核分枝杆菌是否耐药造成影响。
[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.
【学位授予单位】：皖南医学院
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：R52

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