解脲脲原体的多位点序列分型

发布时间：2018-05-21 02:09

本文选题：解脲脲原体 + 多位点序列分型　；参考：《浙江大学》2015年硕士论文

【摘要】：背景及意义解脲脲原体(Ureaplasma, UU)属柔膜纲支原体目支原体科脲原体属,是一种介于细菌与病毒之间的一种能独立生活的最小原核细胞型微生物,它与微小脲原体、生殖脲原体等常常寄居在正常人群生殖道、泌尿道。但也是一种易引起生殖道、泌尿道感染的病原微生物,有大量文献报道,它们会引起男性的前列腺炎、尿道炎,尿结石,肾盂肾炎等,对于女性,还常常是阴道炎、子宫颈炎、盆腔炎、生殖腔道的炎性反应、粘连与阻塞,以及造成不孕不育、流产的病原体之一。曾经我们认为,UU引起的感染是否发生很大程度上取决于人体的抵抗屏障和免疫状态是否正常；但近来有很多文献发现,UU有不同的生物群和亚群,而是否引起感染貌似和他们也有着很大的联系。研究者们已建立了多种针对UU的基因分型方法,包括传统的聚合酶链反应(polymerase chain reaction, PCR),限制性片段长度多态性(restriction fragment length polymorphism, RFLP)等。然而,针对UU特定血清型或亚型与某种特定疾病的关联还未见报道。为了鉴定具有引起特定疾病的UU亚群并研究UU分子流行病学信息,迫切需求新的高分辨率分型方法的建立。本次研究利用基于4个管家基因(ftsH, rpL22, valS, thrS)进行的多位点序列分型(MLST)方法对283株解脲脲原体进行分析,其中包括14种血清型标准菌株及临床样本分离得到的269株解脲脲原体。结果发现MLST序列能够在探索脲原体属分子流行病学及人群构成方面有很高的鉴别能力。目的研究解脲脲原体临床菌株的分子流行病学规律和种群结构,明确遗传背景,为解脲脲原体的感染诊治提供理论参考。方法 269株解脲脲原体临床菌株,其中134株来源于有症状患者,135株来源于无症状人群,与14株标准菌株开展多位点序列分型(MLST)研究,并应用MEGA5与START2软件进行生物信息学分析,以揭示其流行趋势。结果分析283株解脲脲原体发现可分成99种序列型(STs),14种血清型标准菌株分为12种STs,269株临床菌株可分为87种特异性STs。ST1和ST22为主要的STs,分别占有68和70株。eBURST分析表明存在两个克隆复合体(CC1和CC2),并且通过标准连锁指数(IAS)分析显示存在连锁不平衡。系统发育学分析显示解脲脲原体14种血清型标准菌株分为两个种群,此结果与解脲脲原体的物种分群(分为解脲脲原体和微小脲原体两类)高度相符。对269株临床菌株进行生物群分析,CC1中的菌株都是微小脲原体,CC2中的是解脲脲原体,此外,阴道炎、输卵管阻塞及子宫颈炎患者中CC2型感染更为多见。结论解脲脲原体MLST方案具有很高的分辨率,适用于解脲脲原体的分子流行病学和种群结构分析。
[Abstract]:Background and significance Ureaplasma Ureaplasma (UU) belongs to the genus Ureaplasma of Mycoplasma Mycoplasma. Ureaplasma Urealyticum (UU) is a minimal prokaryotic microorganism that can live independently between bacteria and viruses. Reproductive Ureaplasma and so on often sojourn in the normal population genital tract, urinary tract. However, it is also a pathogenic microorganism that can easily cause genital tract, urinary tract infection, which has been reported in a large number of documents. They can cause prostatitis, urethritis, urinary calculi, pyelonephritis and so on in men. For women, they are often vaginitis, cervicitis, etc. Pelvic inflammation, inflammation of the reproductive tract, adhesion and obstruction, and one of the pathogens causing infertility and abortion. We used to think that infection caused by UU depends largely on whether the body's resistance barrier and immune state are normal, but there have been a lot of literature recently found that the UU has different biota and subgroups. Whether or not to cause infection seems to have a strong connection with them. Many genotyping methods for UU have been established, including polymerase chain reactionation (PCR), restriction fragment length polymorphism, RFLP) restriction (restriction fragment length polymorphism, RFLP), and so on. However, the association of UU specific serotypes or subtypes with a specific disease has not been reported. In order to identify the UU subgroups that cause specific diseases and to study the molecular epidemiological information of UU, it is urgent to establish a new high resolution typing method. In this study, 283 strains of Ureaplasma Urealyticum were analyzed by multilocus sequence typing based on four housekeeping genes, ftsH, rpL22, valSST, and 269 strains of Ureaplasma Urealyticum were isolated from 14 serotype standard strains and clinical samples. The results showed that MLST sequence could be used to identify the molecular epidemiology and population composition of Ureaplasma. Purpose To study the molecular epidemiology and population structure of Ureaplasma Urealyticum clinical strains, to clarify the genetic background, and to provide a theoretical reference for the diagnosis and treatment of Ureaplasma Urealyticum infection. Method There were 269 clinical strains of Ureaplasma Urealyticum, of which 134 were from asymptomatic patients and 135 were from asymptomatic population. Multilocus sequence typing was carried out with 14 standard strains, and bioinformatics analysis was carried out with MEGA5 and START2 software. To reveal its trend. Result The analysis of 283 strains of Ureaplasma Urealyticum can be divided into 99 serotypic strains and 14 serotypic standard strains, which can be divided into 12 strains of STS, which can be divided into 87 kinds of specific STs.ST1 and 70 strains of STS. EBURST analysis shows that there are two kinds of STS. The analysis of CC1 and CC2 by the standard linkage index (IASA) showed linkage disequilibrium. Phylogenetic analysis showed that 14 strains of Ureaplasma Urealyticum serotype were divided into two populations, which was highly consistent with the species groups of Ureaplasma Urealyticum and Ureaplasma minimus. Biota analysis of 269 clinical strains of CC1 was carried out. Ureaplasma Urealyticum was found in CC2. In addition, CC2 type infection was more common in patients with vaginitis, tubal obstruction and cervicitis. Conclusion Ureaplasma Urealyticum MLST has high resolution and is suitable for molecular epidemiology and population structure analysis of Ureaplasma Urealyticum.
【学位授予单位】：浙江大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：R446.5

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