本文選題：早產(chǎn)兒　切入點：喂養(yǎng)不耐受　出處：《重慶醫(yī)科大學》2012年碩士論文　論文類型：學位論文

【摘要】：第一部分：PCR-DGGE分析早產(chǎn)兒喂養(yǎng)不耐受腸道菌群 【目的】利用PCR-DGGE技術(shù)分析喂養(yǎng)不耐受早產(chǎn)兒和喂養(yǎng)耐受早產(chǎn)兒（對照組）的大便菌群的結(jié)構(gòu)差異。 【方法】采集15例喂養(yǎng)不耐受早產(chǎn)兒和15例喂養(yǎng)耐受早產(chǎn)兒（對照組）生后第1d，出現(xiàn)喂養(yǎng)不耐受，喂養(yǎng)不耐受恢復后大便標本。提取大便中的細菌混合DNA，再對其16S rDNA的V3可變區(qū)進行PCR擴增，擴增產(chǎn)物經(jīng)DGGE電泳后比較譜帶的異同，找出特征性條帶，，回收測序后確定菌屬。 【結(jié)果】組別相匹配的喂養(yǎng)不耐受組和對照組的DGGE條帶有明顯的差異，多出或缺失幾條明顯的優(yōu)勢條帶，特征性條帶經(jīng)回收測序確定屬于大腸埃希菌、肺炎克雷伯菌、糞腸球菌和乳酸桿菌。 【結(jié)論】喂養(yǎng)不耐受組較對照組而言，腸道的優(yōu)勢菌群發(fā)生了變化。 第二部分Real-Time PCR定量分析早產(chǎn)兒喂養(yǎng)不耐受腸道菌群變化 【目的】觀察早產(chǎn)兒喂養(yǎng)不耐受腸道中大腸埃希菌、糞腸球菌、肺炎克雷伯菌、乳酸桿菌和雙歧桿菌的變化。 【方法】采用實時熒光定量PCR技術(shù)，分別對15例喂養(yǎng)不耐受早產(chǎn)兒和15例喂養(yǎng)耐受早產(chǎn)兒（對照組）生后第1d，出現(xiàn)喂養(yǎng)不耐受，喂養(yǎng)不耐受恢復后大便標本中的大腸埃希菌、肺炎克雷伯菌、糞腸球菌、乳酸桿菌和雙歧桿菌進行定量分析。 【結(jié)果】喂養(yǎng)不耐受組中大腸埃希菌的拷貝數(shù)對數(shù)值（lg copies/g）分別為2.62±0.22、5.47±1.28、3.04±0.70，對照組分別為2.56±0.19、2.82±0.4、2.80±0.39；肺炎克雷伯菌的拷貝數(shù)對數(shù)值（lg copies/g）分別為4.37±0.22、6.56±0.27、4.17±0.27，對照組分別為4.35±0.22、4.19±0.14、4.15±0.25；糞腸球菌的拷貝數(shù)（copies/g）分別為79.17±93.46、42.84±47.57、101.68±43.78，對照組分別為70.16±78.41、740.05±657.71、104.57±38.39；乳酸桿菌的拷貝數(shù)（copies/g）分別為204.03±25.57、326.04±82.42、677.73±559.49，對照組中205.48±16.65、678.79±124.93、663.33±491.57；雙歧桿菌的拷貝數(shù)對數(shù)值（lg copies/g）分別為4.79±0.07、5.27±0.17、5.65±0.25，對照組4.76±0.07、5.57±0.09、5.64±0.15。出現(xiàn)喂養(yǎng)不耐受時，兩組的5種細菌拷貝數(shù)比較差異具有顯著性（P＜0.05），而出現(xiàn)之前和恢復后兩組細菌拷貝數(shù)差異無顯著性（P0.05）。 【結(jié)論】喂養(yǎng)不耐受時大腸埃希菌、肺炎克雷伯菌數(shù)量顯著增高，可能參與喂養(yǎng)不耐受的發(fā)生，而糞腸球菌、乳酸桿菌、雙歧桿菌呈現(xiàn)顯著降低，可能起保護作用。
[Abstract]:Part one: PCR-DGGE analysis of intestinal flora in premature infants with feeding intolerance. [objective] to analyze the differences of fecal flora between preterm infants fed intolerant and those fed with preterm infants (control group) by PCR-DGGE technique. [methods] Fifteen cases of preterm infants with feeding intolerance and 15 cases with feeding tolerance of premature infants (control group) were collected at the first day after birth. Fecal specimens after feeding intolerance recovered, the mixed DNA of bacteria was extracted from feces, and the V3 variable region of 16s rDNA was amplified by PCR. The amplified products were compared with the similarities and differences of the bands by DGGE electrophoresis, and the characteristic bands were found and sequenced to determine the genus of bacteria. [results] there were significant differences in the DGGE bands between the feeding intolerance group and the control group. The DGGE bands were found to be Escherichia coli and Klebsiella pneumoniae, and the characteristic bands were found to belong to Escherichia coli and Klebsiella pneumoniae. Enterococcus faecalis and Lactobacillus. [conclusion] compared with the control group, the dominant flora of the intestinal tract in the feeding intolerance group changed. The second part of Real-Time PCR quantitative analysis of intestinal flora changes of preterm infants with feeding intolerance. [objective] to observe the changes of Escherichia coli, Enterococcus faecalis, Klebsiella pneumoniae, Lactobacillus and Bifidobacterium in feeding intolerance of premature infants. [methods] Fifteen cases of preterm infants fed intolerant and 15 cases fed with preterm infants (control group) were treated with real-time fluorescence quantitative PCR. The first day of feeding intolerance and feeding intolerance of Escherichia coli in fecal specimens after recovery were observed in 15 cases of preterm infants and 15 cases of feeding tolerant preterm infants (control group) at the first day after birth. Klebsiella pneumoniae, Enterococcus faecalis, Lactobacillus and Bifidobacterium were quantitatively analyzed. [results] the logarithmic copy numbers of Escherichia coli in the feeding intolerance group were 2.62 鹵0.225.47 鹵1.283.04 鹵0.70, 2.56 鹵0.192.82 鹵0.4 鹵2.80 鹵0.39, 4.37 鹵0.226.56 鹵0.274.17 鹵0.27, 4.35 鹵0.224.19 鹵0.144.15 鹵0.25, respectively, for Klebsiella pneumoniae, the logarithmic values were 4.37 鹵0.226.56 鹵0.274.17 鹵0.274.17, 4.35 鹵0.224.19 鹵0.144.15 鹵0.25 for Klebsiella pneumoniae, respectively. The copy numbers of Lactobacillus sp. were 79.17 鹵93.46.46 鹵42.84 鹵47.57101.68 鹵43.78, 70.16 鹵78.41740.05 鹵657.71104.57 鹵38.39, and 204.03 鹵25.57326.04 鹵82.42677.73 鹵559.49, 205.48 鹵16.65678.79 鹵124.93663.33 鹵491.57 in the control group, 4.79 鹵0.075.27 鹵0.175.65 鹵0.2565 鹵0.2575 in the control group respectively, and 4.76 鹵0.075.57 鹵0.095.64 鹵0.15. There was a significant difference in the number of copies of the five bacteria between the two groups (P < 0.05), but there was no significant difference in the number of copies between the two groups before and after recovery (P < 0.05). [conclusion] the number of Escherichia coli and Klebsiella pneumoniae increased significantly during feeding intolerance, which may be involved in the development of feeding intolerance, while Enterococcus faecalis, Lactobacillus and Bifidobacterium decreased significantly, which may play a protective role.
【學位授予單位】：重慶醫(yī)科大學
【學位級別】：碩士
【學位授予年份】：2012
【分類號】：R722.6

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