分枝桿菌菌種鑒定及北京家族蛋白質(zhì)組學(xué)初步研究
發(fā)布時(shí)間:2018-02-14 00:38
本文關(guān)鍵詞: 非結(jié)核分枝桿菌 膿腫分枝桿菌 PCR-RFLP 結(jié)核分枝桿菌北京家族 蛋白質(zhì)組學(xué) 出處:《中南大學(xué)》2011年碩士論文 論文類型:學(xué)位論文
【摘要】:目的初步鑒定我國(guó)致病性非結(jié)核分枝桿菌(NTM)、龜-膿腫分枝桿菌復(fù)合群的菌種;探討分枝桿菌快速分子鑒定技術(shù)。初步分析結(jié)核分枝桿菌(MTB)北京家族和卡介苗(BCG)的蛋白質(zhì)組及其差異。 方法用L-J和PNB/TCH培養(yǎng)基對(duì)經(jīng)地方實(shí)驗(yàn)室鑒定為NTM的臨床菌株復(fù)核鑒別后,用hsp65和rpoB PCR-RFLP對(duì)NTM分類鑒定;用細(xì)菌學(xué)、探針雜交和以16s rRNA、16s-23s rRNA、rpoB、hsp65、recA和sodA為靶標(biāo)的多靶位技術(shù)對(duì)龜-膿腫分枝桿菌復(fù)合群進(jìn)行亞種鑒定。蛋白質(zhì)組研究采用雙向凝膠電泳和質(zhì)譜技術(shù)。 結(jié)果復(fù)核鑒定178株菌株中MTB 27株、牛分枝桿菌1株和NTM150株。NTM中胞內(nèi)分枝桿菌72株、膿腫分枝桿菌群27株、戈登分枝桿菌12株、鳥分枝桿菌11株、偶發(fā)分枝桿菌9株、堪薩斯分枝桿菌2株、猿分枝桿菌1株、冬天分枝桿菌1株、M. arupense 1株和未定種14株。膿腫分枝桿菌群中膿腫分枝桿菌19株、馬賽分枝桿菌3株和鑒定出龜-膿腫復(fù)合群新亞種(福建分枝桿菌)5株。分析了北京家族菌株765個(gè)蛋白質(zhì)點(diǎn),發(fā)現(xiàn)了6種特有蛋白質(zhì)和2種表達(dá)明顯增加的蛋白質(zhì)。 結(jié)論中國(guó)NTM肺病病原種類繁多,以胞內(nèi)分枝桿菌群和龜-膿腫分枝桿菌復(fù)合群為主。龜-膿腫分枝桿菌復(fù)合群至少含有三種亞種,并首次發(fā)現(xiàn)了龜-膿腫分枝桿菌復(fù)合群福建分枝桿菌。多靶位分子檢測(cè)技術(shù)聯(lián)合能有效對(duì)NTM進(jìn)行種和亞種鑒定。北京家族與BCG蛋白組間存在明顯差異。
[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.
【學(xué)位授予單位】:中南大學(xué)
【學(xué)位級(jí)別】:碩士
【學(xué)位授予年份】:2011
【分類號(hào)】:R378
【引證文獻(xiàn)】
相關(guān)碩士學(xué)位論文 前1條
1 黃根成;hsp65-、rpoB-PCR-RFLP用于非結(jié)核分枝桿菌菌種鑒定的研究[D];揚(yáng)州大學(xué);2012年
,本文編號(hào):1509473
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