江苏省耐多药结核分枝杆菌分子流行病学研究

发布时间：2018-01-03 01:37

  本文关键词：江苏省耐多药结核分枝杆菌分子流行病学研究　出处：《东南大学》2017年硕士论文　论文类型：学位论文

  更多相关文章： 结核分枝杆菌 耐多药 MIRU-VNTR基因分型 线性探针

【摘要】：目的研究江苏省耐多药结核分枝杆菌(MDR-TB)的分子流行病学特征。(1)建立以结核分枝杆菌分散重复单元一数目可变串联重复序列(mycobacterial interspersed repetitive units-variable number tandem repeats,以下简称 MIRU-VNTR)方法为基础的结核分枝杆菌基因分型系统,通过MIRU-VNTR,分析江苏省不同地区MDR-TB的遗传品系和流行优势菌株,建立MDR-TB基因组多态性数据库。(2)评价MIRU-VNTR分型方法在MDR-TB流行病学中的应用,描述江苏省耐多药结核分枝杆菌的基因型分布及成簇特征,并与非MDR-TB基因型分布及成簇特征进行比较。分析MDR-TB菌株基因型与耐药基因突变间的关联性,了解结核杆菌耐药基因突变可能对菌株的遗传进化产生影响。方法选取2008年江苏省耐药监测项目获得的235株MDR-TB菌株作为研究对象,并随机选取272株非MDR菌株作为对照组,MDR-TB组与非MDR-TB组比例为1:1.2。采用国际通用的标准MIRU-VNTR24位点对纳入的全部结核分枝杆菌进行基因型检测,并采用HAIN线性探针技术对MDR-TB利福平和异烟肼的耐药突变位点进行检测。MIRU-VNTR数据通过网站http://www.miru-vntrplus.org进行聚类分析,并采用非参数检验比较两组的统计学意义,并对成簇菌株的耐药突变信息进行比较分析。结果对MDR-TB和非MDR-TB的MIRU-VNTR的基因多态性进行比较,得出235株MDR分为206个基因型,其中185个独特性,有17个基因型有两株,3个基因型有3株,1个基因型有7株。成簇率为21.3%(50/235)。272株非MDR分为229个基因型,其中210个独特性,11个基因型含2株,4个基因型含3株,2个基因型含4株,1个基因型含7株,1个基因型含13株,成簇率为22.8%(62/272),比较MDR菌株与非MDR菌株两组的成簇率,无统计学差异(X2=0.169,p值为0.681),提示MDR与非MDR在聚集性上并没有明显差异。MIRU-VNTR24位点对235株MDR-TB和272株非MDR-TB进行基因检测,采用非参数检验比较发现两组在24个位点的分布上并无统计学意义,计算每个位点的分辨率指数,并挑选了分辨率指数大于0.3的13个位点,另外两组比较P值小于0.1的一个位点,共14的位点,分别为:ETRA、ETRE、MIRU10、MIRU23、MIRU26、MIRU39、MIRU40、Mtub04、Mtub21、Mtub30、Mtub39、Oub11b、Qub26、Qub4156c。比较两组 24 个位点与 14 个位点的分辨率分别为 MDR-24 分辨率为 0.9982,非 MDR-24 为 0.9964,MDR-14 为 0.9958,非 MDR-14为0.9967。筛选的14个位点的分辨能力与24个位点无显著差异。采用HAIN线性探针技术快速检测方法对235株MDR进行了利福平耐药基因(rpoB)和异烟肼耐药基因(katG、inhA)的突变情况检测,获得MDR-TB耐药基因突变类型,对成簇的结核分枝杆菌耐药突变位点进行进一步的分析发现成簇的结核分枝杆菌其耐药突变位点并不相似,提示菌株是否成簇与其耐药突变无直接关联。结论江苏省MDR-TB与非MDR-TB在基因多态性无统计学差异。结核分枝杆菌成簇性与耐药基因无相关性。现行的普通结核病防治策略可适用于耐多药结核病的防治。
[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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