北京地区结核分枝杆菌基因分型及与耐药性关系的研究

发布时间：2017-12-27 11:15

本文关键词：北京地区结核分枝杆菌基因分型及与耐药性关系的研究　出处：《北京市结核病胸部肿瘤研究所》2012年博士论文　论文类型：学位论文

【摘要】：目的了解北京地区679株分枝杆菌临床分离株的基因型分布、主要流行特点,比较465株分枝杆菌间隔区寡核苷酸分型(Spoligotyping)和散在分布重复单位-数目可变串联重复序列(MIRU-VNTR)两种基因型方法试验结果的异同,分析北京基因型与耐药性的相互关系,从而探讨北京地区结核分枝杆菌的分子流行特点,分析结核分枝杆菌的耐药性特点及其同分子流行病学的联系,并评价两种基因分型方法。方法对收集的北京市679株分枝杆菌临床分离株进行Spoligotyping基因分型研究,其中的465株分枝杆菌同时运用MIRU-VNTR方法进行基因分型研究。用NatureEdge软件和BioNumerics软件将分型结果进行聚类分析。应用比例法药物敏感试验对268株结核分枝杆菌进行检测,最后将药敏结果结合流行病学资料进行分析。采用χ2检验比较不同组间结果的差别。结果在分离培养的679株菌株中,具有特异Spoligotyping指纹图谱的北京基因型菌株在北京地区分布达82.6%(561/679),为北京地区2008年主要流行的基因型菌株。未接种过卡介苗(BCG)的患者中北京基因型菌株占83.5%(240/269),接种过BCG的患者中北京基因型菌株占81.7%(384/470),两者差异无统计学意义。十种药物耐药性由高到低依次为丁胺卡那霉素(61.76%)、利福平(54.65%)、乙硫异烟胺(44.16%)、利福布汀(39.71%)、异烟肼(23.37%)、链霉素(22.22%)、左氧氟沙星(18.87%)、对氨基水杨酸(8.70%)、卷曲霉素(6.52%)、乙胺丁醇(4.71%)。北京基因型菌株耐药率为86.11%(186/216),低于非北京基因型菌株的耐药率93.18%(41/44),但二者差异也无统计学意义。Spoligotyping分型方法显示成簇菌株占所有菌株的93.93%(437/465),分73个基因型,MIRU-VNTR分型方法显示成簇菌株占所有菌株的22.37%(104/465),398个基因型。本地户籍成为菌株成簇的一个危险因素,OR值为1.90。VNTR方法对465株分枝杆菌的分辨率指数为0.9576。对北京基因型的分辨率指数为0.9547,同样显示了很高的分辨率指数。而Spoligotyping分型方法对465株分枝杆菌的分辨率指数0.4131,对北京基因型的分辨率指数为0.1430,低于MIRU-VNTR方法。不同的菌株和同一菌株在不同位点的MIRU-VNTR DNA指纹图谱呈现出多态性,各位点呈现的等位基因多态性有差异,多态性最高的位点是QUB-11b,多态性最低的位点为QUB-4156c。对北京基因型位点多态性分析也显示了相同的最高位点和最低位点。结论北京基因型菌株2008年在北京地区有广泛分布,为占绝对优势的菌株；北京基因型菌株与BCG接种无关,与一线、二线抗结核药物耐药无关；结核病患者中有部分是由于近期传播而引起的。MIRU-VNTR基因分型方法能够对Spoligotyping分型结果继续分型,分辨率指数要高于Spoligotyping分型方法。MIRU-VNTR分型方法对所有基因型和对北京基因型的分辨指数差别不大。Spoligotyping分型方法对北京基因型的分辨率指数较低。
[Abstract]:Objective to understand the Beijing region of 679 strains of genotype of Mycobacterium tuberculosis strains distribution, epidemic characteristics, 465 strains of Mycobacterium type spacer oligonucleotide (Spoligotyping) and scattered distribution of variable number of tandem repeat repeat units (MIRU-VNTR) between the two genotypes of test results, analysis of the relationship between Beijing genotype with the resistance, so as to explore the molecular epidemiological characteristics of Mycobacterium tuberculosis in Beijing area, with the analysis of resistance characteristics and molecular epidemiology of Mycobacterium tuberculosis contact, and evaluate the two genotyping methods. Methods a total of 679 Mycobacterium clinical isolates from Beijing were analyzed for Spoligotyping genotyping. 465 Mycobacterium strains were genotyping by MIRU-VNTR. NatureEdge software and BioNumerics software were used to cluster analysis. 268 strains of Mycobacterium tuberculosis were detected by using the proportion method of drug sensitivity test. Finally, the results of drug sensitivity were analyzed with epidemiological data. The difference between the results of different groups was compared with the x 2 test. Results among the 679 strains isolated and cultured, the Beijing genotype Spoligotyping with specific 561/679 fingerprints distributed 82.6% (561/679) in Beijing area, which was the most popular genotype in Beijing area. Among the patients who had not been vaccinated with Bacillus Calmette Guerin vaccine (BCG), the genotype of Beijing accounted for 83.5% (240/269), while the genotype of Beijing genotype accounted for 81.7% (384/470), and the difference between the two was not statistically significant (BCG). Ten kinds of drug resistance from high to low were amikacin (61.76%), Li Fuping (54.65%), ethionamide (44.16%), Leigh Fob Dean (39.71%), (23.37%) isoniazid and streptomycin (22.22%), levofloxacin (18.87%), salicylic acid (8.70%), capreomycin (6.52%), ethambutol (4.71%). The resistance rate of Beijing genotype was 86.11% (186/216), and the resistance rate of the non Beijing genotype was 93.18% (41/44), but the difference between the two strains was not statistically significant. The Spoligotyping typing method showed that the cluster strains accounted for 93.93% (437/465) of all strains, which were divided into 73 genotypes. The MIRU-VNTR typing method showed that the cluster strains accounted for 22.37% (104/465) and 398 genotypes of all strains. Local domicile has become a risk factor for the cluster of strains, with a OR value of 1.90. The resolution index of the VNTR method for 465 Mycobacterium strains was 0.9576. The resolution index of the Beijing genotypes was 0.9547, which also showed a high resolution index. The resolution index of Spoligotyping typing for 465 strains of Mycobacterium was 0.4131, and the resolution index of Beijing genotypes was 0.1430, which was lower than that of MIRU-VNTR. The MIRU-VNTR DNA fingerprints of different strains and the same strain at different loci showed polymorphism. The alleles polymorphism at each loci was different. The most polymorphic locus was QUB-11b, and the lowest polymorphism locus was QUB-4156c. The polymorphism analysis of the genotypic loci in Beijing also showed the same highest site and the lowest point. Conclusion Beijing genotype strains were widely distributed in Beijing area in 2008. They are the most dominant strains. The genotype of Beijing has nothing to do with BCG vaccination. It is not related to the resistance of first-line and second-line anti tuberculosis drugs. Some of the tuberculosis patients are caused by recent transmission. The MIRU-VNTR genotyping method can continue to be typed for the results of Spoligotyping typing, and the resolution index is higher than that of the Spoligotyping typing. MIRU-VNTR typing has little difference in the resolution index of all genotypes and Beijing genotypes. The resolution index of the genotyping of the Beijing genotypes was lower than that of the Beijing genotyping.
【学位授予单位】：北京市结核病胸部肿瘤研究所
【学位级别】：博士
【学位授予年份】：2012
【分类号】：R378.911

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