分枝杆菌菌种鉴定及北京家族蛋白质组学初步研究

发布时间：2018-02-14 00:38

本文关键词： 非结核分枝杆菌脓肿分枝杆菌 PCR-RFLP 结核分枝杆菌北京家族蛋白质组学　出处：《中南大学》2011年硕士论文　论文类型：学位论文

【摘要】：目的初步鉴定我国致病性非结核分枝杆菌(NTM)、龟-脓肿分枝杆菌复合群的菌种；探讨分枝杆菌快速分子鉴定技术。初步分析结核分枝杆菌(MTB)北京家族和卡介苗(BCG)的蛋白质组及其差异。方法用L-J和PNB/TCH培养基对经地方实验室鉴定为NTM的临床菌株复核鉴别后,用hsp65和rpoB PCR-RFLP对NTM分类鉴定；用细菌学、探针杂交和以16s rRNA、16s-23s rRNA、rpoB、hsp65、recA和sodA为靶标的多靶位技术对龟-脓肿分枝杆菌复合群进行亚种鉴定。蛋白质组研究采用双向凝胶电泳和质谱技术。结果复核鉴定178株菌株中MTB 27株、牛分枝杆菌1株和NTM150株。NTM中胞内分枝杆菌72株、脓肿分枝杆菌群27株、戈登分枝杆菌12株、鸟分枝杆菌11株、偶发分枝杆菌9株、堪萨斯分枝杆菌2株、猿分枝杆菌1株、冬天分枝杆菌1株、M. arupense 1株和未定种14株。脓肿分枝杆菌群中脓肿分枝杆菌19株、马赛分枝杆菌3株和鉴定出龟-脓肿复合群新亚种(福建分枝杆菌)5株。分析了北京家族菌株765个蛋白质点,发现了6种特有蛋白质和2种表达明显增加的蛋白质。结论中国NTM肺病病原种类繁多,以胞内分枝杆菌群和龟-脓肿分枝杆菌复合群为主。龟-脓肿分枝杆菌复合群至少含有三种亚种,并首次发现了龟-脓肿分枝杆菌复合群福建分枝杆菌。多靶位分子检测技术联合能有效对NTM进行种和亚种鉴定。北京家族与BCG蛋白组间存在明显差异。
[Abstract]:Objective to identify the pathogenic nontuberculous mycobacteria (NTMN) and mycobacterium tortoise-abscess (MBS) in China. To study the rapid molecular identification technique of Mycobacterium tuberculosis, the proteome and its difference between the Beijing family of Mycobacterium tuberculosis (MTB) and the BCG (Bacillus Calmette Calmette Calmette Guerin) were analyzed. Methods L-J and PNB/TCH culture medium were used to identify the clinical strains identified as NTM by local laboratory, hsp65 and rpoB PCR-RFLP were used to classify and identify NTM, bacteriology, bacteriology, and bacteriology were used to identify NTM. Probe hybridization and multi-target techniques targeting 16s-23s rRNA-rRNA-rRNA-rRNA-rRNA-rRNA-rRNA-rRNA-rRNA-rRNA-rRNA-rRNA-rRNA-rRNA-rRNA-rRNA-rRNA-rRNA-rRNA-rRNA-rRNA-rRNA-rRNA-rRNA-rRNA-rRNA-rRNA-rRNA-rRNA-rRNA-rRNA@@. Results there were 27 strains of MTB, 1 strain of Mycobacterium bovis, 72 strains of endocytobacillus in NTM150 strain, 27 strains of mycobacterium abscess, 12 strains of Mycobacterium Gordon, 11 strains of Mycobacterium bird, 9 strains of Mycobacterium chance. Two strains of Mycobacterium Kansas, one strain of Mycobacterium ape, one strain of M. arupense and 14 strains of undetermined species. Three strains of Mycobacterium Maxima and 5 new subspecies of Turtle abscess complex group (Mycobacterium Fujian) were identified. 765 protein spots of Beijing family strain were analyzed, and 6 specific proteins and 2 proteins with significantly increased expression were found. Conclusion there are many pathogens of NTM's pulmonary disease in China, which are mainly composed of intracellular mycobacterium and mycobacterium tortoise and abscess, and there are at least three subspecies in the complex group of mycobacterium tortoise and abscess. The combination of multi-target molecular detection and multitarget molecular detection can effectively identify the species and subspecies of NTM. There are significant differences between Beijing family and BCG protein group.
【学位授予单位】：中南大学
【学位级别】：硕士
【学位授予年份】：2011
【分类号】：R378

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