造血干细胞移植病人与肠道微生态的关系

发布时间：2018-03-11 17:16

本文选题：干细胞移植　切入点：肠道菌群　出处：《复旦大学》2014年硕士论文　论文类型：学位论文

【摘要】：[目的]本研究通过对干细胞移植患者与正常对照者之间粪便菌群种类及特异菌群的定量研究,了解干细胞移植患者肠道菌群结构差异及数量的变化,并探讨干细胞移植患者是否出现肠道细菌结构紊乱从而引起院内感染。[方法]本研究为前瞻性,观察性研究。根据入选标准,纳入复旦大学附属华东医院血液科进行干细胞移植的患者,收集患者干细胞移植前7-10天、干细胞移植后7-10天,干细胞移植后发生感染时的粪便标本。另外收集20个正常人的粪便标本作为对照。根据细菌的16SrDNA基因序列设计大肠埃希菌、粪肠球菌属、乳酸杆菌属、及拟杆菌属的引物,运用实时荧光定量-聚合酶链反应法(RT-PCR)定量检测两组患者中这四种细菌的数量变化。[结果]经RT-PCR对大肠埃希菌、肠球菌属、乳酸杆菌属、拟杆菌属定量检测后发现：1.HSCT患者3组相比：造血干细胞移植后发生感染时、干细胞移植前7-10天及干细胞移植后7-10天相比,粪便中3种细菌(大肠埃希菌、肠球菌属、和乳酸杆菌属)数量增多,其变化量分别为大肠埃希菌8.66±2.65 vs 8.55±1.1l vs 7.97±0.49, p=0.735；肠球菌属7.47±1.75 vs 6.70±1.76 vs 7.09±1.54, p=0.14:乳酸杆菌9.03±0.29 vs 7.57±0.28 vs 8.69±0.19 p=0.009.而在造血干细胞移植后7-10天,患者肠道中拟杆菌属数量增加,与造血干细胞移植后发生感染时及干细胞移植前7-10天相比,分别为6.74±0.19 vs 6.58±0.28 vs 6.67±0.24, p=0.964。2.造血干细胞移植后发生感染时肠道细菌数量较对照组增加,分别为大肠埃希菌8.66±2.65 vs 8.51±0.54,p=0.795,肠球菌属7.47±1.75 vs7.11±1.06,p=0.000,乳酸杆菌属9.03±0.29 vs 8.67±0.43,p=0.72,拟杆菌属6.67±0.24 vs 6.66±0.28,p=0.243。与对照组相比,造血干细胞移植前7-10天患者肠道中大肠埃希菌及粪球菌属数量下降,分别为7.97±0.49vs 8.51±0.54,p=0.417；7.09±1.54 vs 7.11±1.06,p=0.580。而乳酸杆菌属及拟杆菌属数量增多,分别为8.69±0.19 vs 8.67±0.43,p=0.954；6.74±0.19vs 6.66±0.28,p=0.85.3. HSCT患者3个采样点比较,只有乳酸杆菌属数量有显著差异(p=0.009),其他菌种之间无显著差异(p0.05)。而造血干细胞移植后发生感染时与对照组相比,4种细菌都有所增加,分别为大肠埃希菌8.66±2.65 vs 8.51±0.54,肠球菌属7.47±1.75 vs 7.11±1.06,乳酸杆菌属9.03±0.29 vs 8.67±0.43和拟杆菌属6.67±0.24 vs6.66±0.28),但只有肠球菌属的差异具有统计学意义(p=0.000)。4.造血干细胞移植前7-10天大便菌群数量总体较对照组有所下降,但只乳酸杆菌属的差异具有统计学意义(p=0.002)。[结论]1.与对照组相比,造血干细胞移植后发生感染时肠道细菌明显增多。当造血干细胞移植后发生感染时肠道细菌结构发生改变。2.当造血干细胞移植后发生感染时肠道细菌结构发生改变,主要为肠道兼性厌氧细菌,其中以乳酸杆菌属及大肠埃希菌最为显著。3. Real-time PCR方法检测粪便中细菌DNA能反映肠道细菌改变。
[Abstract]:[objective] to investigate the difference and quantity of intestinal flora in stem cell transplantation patients by quantitative analysis of fecal microflora and specific microflora between stem cell transplantation patients and normal controls. And to explore whether stem cell transplantation patients have intestinal bacterial structure disorder and cause nosocomial infection. [methods] this study is a prospective, observational study, according to the inclusion criteria, Patients admitted to the Department of Hematology of Huadong Hospital affiliated to Fudan University for stem cell transplantation were collected 7-10 days before transplantation and 7-10 days after stem cell transplantation. Fecal specimens from infected stem cells were collected from 20 normal controls. Primers were designed for Escherichia coli, Enterococcus faecalis, Lactobacillus and Bacteroides according to the 16s rDNA gene sequence of bacteria. Real-time fluorescence quantification-polymerase chain reaction (RT-PCR) was used to detect the quantitative changes of the four bacteria in the two groups. [results] RT-PCR was used to detect Escherichia coli, Enterococcus and Lactobacillus. The results of quantitative analysis of Bacteroides showed that: 1. Three kinds of bacteria (Escherichia coli, Enterococcus) were found in feces when infection occurred after hematopoietic stem cell transplantation, 7-10 days before stem cell transplantation and 7-10 days after stem cell transplantation. The number of Escherichia coli (8.66 卤2.65 vs 8.55 卤1.41 vs 7.97 卤0.49, pn0.735), Enterococcus 7.47 卤1.75 vs 6.70 卤1.76 vs 7.09 卤1.54, p0.14: Lactobacillus 9.03 卤0.29 vs 7.57 卤0.28 vs 8.69 卤0.19 pc0.0099.The number of bacilli in intestinal tract increased 7-10 days after hematopoietic stem cell transplantation. Compared with infection after hematopoietic stem cell transplantation and 7 to 10 days before stem cell transplantation, the number of intestinal bacteria was increased after hematopoietic stem cell transplantation compared with that in control group, which was 6.74 卤0.19 vs 6.58 卤0.28 vs 6.67 卤0.24, and 0.964.2, respectively. The number of Escherichia coli was 8.66 卤2.65 vs 8.51 卤0.54p 0.795. Enterococcus was 7.47 卤1.75 vs7.11 卤1.06p0.000. Lactobacillus was 9.03 卤0.29 vs 8.67 卤0.43p0.72, bacillus was 6.67 卤0.24 vs 6.66 卤0.28p0.243. Compared with the control group, the number of Escherichia coli and S. faecalis decreased 7-10 days before hematopoietic stem cell transplantation. It was 7.97 卤0.49 vs 8.51 卤0.54 卤0.4177.09 卤1.54 vs 7.11 卤1.06 p0. 580.The number of Lactobacillus and Bacteroides increased 8.69 卤0.19 vs 8.67 卤0.43p0. 74 卤0. 19 vs 6. 66 卤0. 28p0. 85.3. HSCT. Only the number of Lactobacillus was significantly different from that of the control group, but there was no significant difference among other strains. However, the number of four bacteria increased after hematopoietic stem cell transplantation (HSCT) compared with the control group. Escherichia coli 8.66 卤2.65 vs 8.51 卤0.54, Enterococcus 7.47 卤1.75 vs 7.11 卤1.06, Lactobacillus 9.03 卤0.29 vs 8.67 卤0.43 and Bacteroides 6.67 卤0.24 vs6.66 卤0.28, respectively, but only Enterococcus showed significant difference in the number of feces from 7 to 10 days before hematopoietic stem cell transplantation. The control group decreased, But the difference of Lactobacillus is statistically significant. [conclusion] 1.Compared with the control group, After hematopoietic stem cell transplantation (HSCT), the intestinal bacterial structure was changed. 2. When the infection occurred after hematopoietic stem cell transplantation (HSCT), the intestinal bacterial structure changed, and the infection occurred after hematopoietic stem cell transplantation (HSCT). The main intestinal facultative anaerobic bacteria were Lactobacillus and Escherichia coli. The detection of bacterial DNA in feces by Real-time PCR could reflect the change of intestinal bacteria.
【学位授予单位】：复旦大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R574

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