结核病诊断及耐药检测分子标识物的发现

发布时间：2019-03-07 17:57

【摘要】：目前，结核病（TB）仍然危害着人类的健康。迫切需要更加有效的诊断结核病的生物标志物，可以在结核病预防、诊断和治疗中用于区分健康个体和潜伏感染人群。本实验通过Solexa测序手段，研究活动性肺结核、潜伏性感染者和接种或未接种卡介苗的未发病人血清中microRNA的表达。结果显示，与其它三组相比，结核病患者的血清有24个microRNA的表达量显著上调和6个microRNA的表达量显著下调。将差异表达的microRNA进行microRNA-基因网络分析和影响途径的分析，结果表明，microRNA很可能通过影响宿主免疫系统的调节使之患上结核病。qRT-PCR实验验证了hsa-miR-196b和hsa-miR-376c在结核病患者血清中的差异表达，可能是潜在的结核病分子标志物，有用作结核病血清检测的潜力。本研究中得到的microRNA差异表达图谱为结核病诊断技术的发展提供了良好的基础，也可能用作鉴别卡介苗接种者和潜伏感染者的候选microRNA。此外，由于多药耐药（MDR）和广泛抗药性（XDR）结核分枝杆菌的出现，使结核病在世界范围内出现越来越难以治愈的情况。目前，结核分枝杆菌的耐药机制还不是很了解，只是掌握了部分结核分枝杆菌耐药性的遗传特点。本次研究中，通过161株结核分枝杆菌的测序和分析，新发现了72个基因，28个基因间隔区与结核分枝杆菌的耐药性有很强的相关性。同时，通过dN/dS的比率分析结果提示，，本研究发现的耐药相关基因很可能包含所有在本研究使用的所有菌株中与药物压力相关的非同义SNPs，这些非同义SNPs所在的基因群展示了一个近乎完整的药物抗性相关的基因和抗生素的图谱。本研究结果还表明，结核分枝杆菌耐药性的遗传机制比以前预想的更为复杂，这些发现为阐明未知的耐药机制奠定坚实的实验基础。
[Abstract]:At present, tuberculosis (TB) is still harmful to human health. There is an urgent need for more effective biomarkers for the diagnosis of tuberculosis, which can be used to distinguish healthy individuals from latent infections in tuberculosis prevention, diagnosis and treatment. In this study, Solexa sequencing was used to study the expression of microRNA in sera of active pulmonary tuberculosis, latent infection and non-infected patients inoculated or not vaccinated with BCG. The results showed that compared with the other three groups, the expression of 24 microRNA and 6 microRNA were significantly up-regulated and down-regulated in TB patients. The differentially expressed microRNA was analyzed by microRNA- gene network analysis and influence pathway analysis. The results showed that: 1. MicroRNA is likely to cause tuberculosis by affecting the immune system of the host. The qRT-PCR experiment verified the differential expression of hsa-miR-196b and hsa-miR-376c in the serum of TB patients, which may be a potential molecular marker of TB. It has the potential to be used as a serum test for tuberculosis. The differential expression map of microRNA obtained in this study provides a good basis for the development of tuberculosis diagnosis techniques, and may also be used as a candidate microRNA. for the identification of BCG vaccinators and latent infections. In addition, due to the emergence of multidrug-resistant (MDR) and extensively drug-resistant (XDR) Mycobacterium tuberculosis, tuberculosis is becoming more and more difficult to cure in the world. At present, the mechanism of drug resistance of Mycobacterium tuberculosis is not well understood, but the genetic characteristics of resistance of some Mycobacterium tuberculosis have been grasped. In this study, through the sequencing and analysis of 161 strains of Mycobacterium tuberculosis, 72 new genes were found, and 28 spacers had strong correlation with the drug resistance of Mycobacterium tuberculosis. At the same time, the results of dN/dS ratio analysis suggest that the drug resistance-related genes in this study may contain all non-synonymous SNPs, associated with drug stress in all strains used in this study. These non-synonymous SNPs gene groups present a near-complete map of drug resistance-related genes and antibiotics. The results also show that the genetic mechanism of drug resistance of Mycobacterium tuberculosis is more complex than previously expected. These findings provide a solid experimental basis for elucidating the unknown mechanism of drug resistance.
【学位授予单位】：中国科学院研究生院（近代物理研究所）
【学位级别】：博士
【学位授予年份】：2014
【分类号】：R52

【共引文献】