本文關(guān)鍵詞：江蘇省耐多藥結(jié)核分枝桿菌分子流行病學(xué)研究　出處：《東南大學(xué)》2017年碩士論文　論文類型：學(xué)位論文

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【摘要】：目的研究江蘇省耐多藥結(jié)核分枝桿菌(MDR-TB)的分子流行病學(xué)特征。(1)建立以結(jié)核分枝桿菌分散重復(fù)單元一數(shù)目可變串聯(lián)重復(fù)序列(mycobacterial interspersed repetitive units-variable number tandem repeats,以下簡稱 MIRU-VNTR)方法為基礎(chǔ)的結(jié)核分枝桿菌基因分型系統(tǒng),通過MIRU-VNTR,分析江蘇省不同地區(qū)MDR-TB的遺傳品系和流行優(yōu)勢菌株,建立MDR-TB基因組多態(tài)性數(shù)據(jù)庫。(2)評價MIRU-VNTR分型方法在MDR-TB流行病學(xué)中的應(yīng)用,描述江蘇省耐多藥結(jié)核分枝桿菌的基因型分布及成簇特征,并與非MDR-TB基因型分布及成簇特征進行比較。分析MDR-TB菌株基因型與耐藥基因突變間的關(guān)聯(lián)性,了解結(jié)核桿菌耐藥基因突變可能對菌株的遺傳進化產(chǎn)生影響。方法選取2008年江蘇省耐藥監(jiān)測項目獲得的235株MDR-TB菌株作為研究對象,并隨機選取272株非MDR菌株作為對照組,MDR-TB組與非MDR-TB組比例為1:1.2。采用國際通用的標準MIRU-VNTR24位點對納入的全部結(jié)核分枝桿菌進行基因型檢測,并采用HAIN線性探針技術(shù)對MDR-TB利福平和異煙肼的耐藥突變位點進行檢測。MIRU-VNTR數(shù)據(jù)通過網(wǎng)站http://www.miru-vntrplus.org進行聚類分析,并采用非參數(shù)檢驗比較兩組的統(tǒng)計學(xué)意義,并對成簇菌株的耐藥突變信息進行比較分析。結(jié)果對MDR-TB和非MDR-TB的MIRU-VNTR的基因多態(tài)性進行比較,得出235株MDR分為206個基因型,其中185個獨特性,有17個基因型有兩株,3個基因型有3株,1個基因型有7株。成簇率為21.3%(50/235)。272株非MDR分為229個基因型,其中210個獨特性,11個基因型含2株,4個基因型含3株,2個基因型含4株,1個基因型含7株,1個基因型含13株,成簇率為22.8%(62/272),比較MDR菌株與非MDR菌株兩組的成簇率,無統(tǒng)計學(xué)差異(X2=0.169,p值為0.681),提示MDR與非MDR在聚集性上并沒有明顯差異。MIRU-VNTR24位點對235株MDR-TB和272株非MDR-TB進行基因檢測,采用非參數(shù)檢驗比較發(fā)現(xiàn)兩組在24個位點的分布上并無統(tǒng)計學(xué)意義,計算每個位點的分辨率指數(shù),并挑選了分辨率指數(shù)大于0.3的13個位點,另外兩組比較P值小于0.1的一個位點,共14的位點,分別為:ETRA、ETRE、MIRU10、MIRU23、MIRU26、MIRU39、MIRU40、Mtub04、Mtub21、Mtub30、Mtub39、Oub11b、Qub26、Qub4156c。比較兩組 24 個位點與 14 個位點的分辨率分別為 MDR-24 分辨率為 0.9982,非 MDR-24 為 0.9964,MDR-14 為 0.9958,非 MDR-14為0.9967。篩選的14個位點的分辨能力與24個位點無顯著差異。采用HAIN線性探針技術(shù)快速檢測方法對235株MDR進行了利福平耐藥基因(rpoB)和異煙肼耐藥基因(katG、inhA)的突變情況檢測,獲得MDR-TB耐藥基因突變類型,對成簇的結(jié)核分枝桿菌耐藥突變位點進行進一步的分析發(fā)現(xiàn)成簇的結(jié)核分枝桿菌其耐藥突變位點并不相似,提示菌株是否成簇與其耐藥突變無直接關(guān)聯(lián)。結(jié)論江蘇省MDR-TB與非MDR-TB在基因多態(tài)性無統(tǒng)計學(xué)差異。結(jié)核分枝桿菌成簇性與耐藥基因無相關(guān)性�，F(xiàn)行的普通結(jié)核病防治策略可適用于耐多藥結(jié)核病的防治。
[Abstract]:Objective to study the Jiangsu province multidrug resistant Mycobacterium tuberculosis (MDR-TB) molecular epidemiological features. (1) to establish Mycobacterium tuberculosis dispersed repeat unit of a variable number of tandem repeats (mycobacterial interspersed repetitive Units-Variable number tandem repeats, hereinafter referred to as MIRU-VNTR) method of Mycobacterium tuberculosis gene based typing system, through the MIRU-VNTR, genetic analysis of MDR-TB strains in different regions of Jiangsu province and the dominant strain, establish MDR-TB genomic polymorphism database. (2) to evaluate the application of MIRU-VNTR classification method in MDR-TB epidemiology, describe the genotype distribution and clustering features of Jiangsu province multidrug resistant Mycobacterium tuberculosis, and compare with the genotype distribution and clustering characteristics MDR-TB. Correlation analysis of MDR-TB strains of genotype and drug resistance mutations among the Mycobacterium tuberculosis resistant gene mutation may understand The influence of genetic strains. Methods from 2008 Jiangsu province drug resistance monitoring project to obtain the 235 MDR-TB strains as the research object, and randomly selected 272 strains of non MDR strains as control group, the proportion of MDR-TB group and non MDR-TB group 1:1.2. using standard MIRU-VNTR24 site international common to all the included genotype of Mycobacterium tuberculosis detection, and using HAIN linear probe technique on MDR-TB of rifampin and isoniazid resistance mutations by cluster analysis of site http://www.miru-vntrplus.org was detected with.MIRU-VNTR data, and test the statistical significance between the two groups by non parametric, and resistance to clusters of mutant information were analyzed. Results the gene polymorphism of MDR-TB and non MDR-TB MIRU-VNTR the comparison, obtained 235 MDR strains into 206 genotypes, including 185 unique, 17 gene type 鏈変袱鏍，

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