本文關(guān)鍵詞： 腸道菌群動態(tài)分析 巴黎鏈球菌 全基因組 毒力島　出處：《中國疾病預(yù)防控制中心》2012年博士論文　論文類型：學(xué)位論文

【摘要】：通過對連續(xù)采集的糞便標(biāo)本中16S rDNA的擴增、克隆和序列分析,我們可以了解腹瀉兒童腸道菌群的動態(tài)變化,這是常規(guī)分離培養(yǎng)方法所不能實現(xiàn)的,同時為研究未知的腹瀉病原菌提供了可能。本文使用細菌16S rDNA序列分析方法,對33名感染性腹瀉兒童糞便標(biāo)本的細菌菌群進行了研究。通過常見腹瀉病原菌的篩查,11名患兒被診斷為致瀉性大腸桿菌或志賀菌感染,其他22名腹瀉兒童被診斷為未知病原感染性腹瀉。在這22名腹瀉兒童中,有12名只采集到入院首日的標(biāo)本,另外10名采集到三份連續(xù)標(biāo)本,分別是入院首日、病情恢復(fù)中和恢復(fù)后或者是出院后的糞便標(biāo)本。在10名患兒入院首日的糞便標(biāo)本中,大腸桿菌和克雷伯菌占優(yōu)勢的有2份,唾液鏈球菌為優(yōu)勢的有2份,牛鏈球菌群及鏈球菌屬某種占優(yōu)勢各有3份。通過對連續(xù)采集的糞便標(biāo)本分析發(fā)現(xiàn),10例患兒中有8例其入院時標(biāo)本的優(yōu)勢菌種隨著治療的過程比例降低,其中以牛鏈球菌群為入院時優(yōu)勢菌種的三名腹瀉兒童,隨著病情的好轉(zhuǎn)其糞便中的牛鏈球菌群序列消失不見。 我們對16S rDNA提示的優(yōu)勢菌群進行分離培養(yǎng),并對分離到的菌株進行生化反應(yīng)和16S rDNA近似全長序列測定。通過分析發(fā)現(xiàn),3份牛鏈球菌群為優(yōu)勢的標(biāo)本分別是巴黎鏈球菌1份,解沒食子酸鏈球菌巴氏亞種2份。我們對分離的巴黎鏈球菌進行全基因組序列分析,發(fā)現(xiàn)巴黎鏈球菌基因組上存在20個基因組島。其中基因組島7(編碼糖基轉(zhuǎn)移酶)與基因組島18(編碼菌毛相關(guān)基因)被認(rèn)為是可能的毒力島。同時發(fā)現(xiàn)其他8個可能的致病基因。 我們首次報道巴黎鏈球菌是部分未知病原感染性腹瀉兒童腸道菌群的優(yōu)勢菌種,其比例隨著治療過程下降。我們對分離的巴黎鏈球菌進行全基因組分析,發(fā)現(xiàn)了可能的毒力島和致病基因。我們分離了腹瀉病人及動物宿主糞便中牛鏈球菌群菌株,并對其進行了分子流行病的研究。
[Abstract]:By amplification, cloning and sequence analysis of 16s rDNA in fecal samples collected continuously, we can understand the dynamic changes of intestinal flora in children with diarrhea, which can not be realized by conventional isolation and culture methods. At the same time, it is possible to study the unknown diarrhea pathogens. The 16s rDNA sequence analysis method of bacteria was used in this paper. The bacterial flora in fecal specimens of 33 children with infectious diarrhea was studied. 11 children were diagnosed as cathartic Escherichia coli or Shigella infection through screening of common diarrhea pathogens. The other 22 children with diarrhea were diagnosed with infectious diarrhea of unknown etiology. Of the 22 children with diarrhoea, 12 were collected only on the first day of admission and 10 received three consecutive specimens on the first day of admission. Stool samples from the patients recovered and recovered or discharged from hospital. Of the 10 children, 2 were Escherichia coli and Klebsiella, and 2 were Streptococcus saliva, out of the 10 children who were hospitalized on the first day of hospitalization, 2 were Escherichia coli and 2 were Klebsiella. There were 3 strains of Streptococcus Bovis and 3 strains of Streptococcus Bovis respectively. By analyzing the fecal samples collected continuously, it was found that 8 out of 10 cases had decreased the proportion of dominant bacteria in the samples when they were admitted to hospital. Among them, three diarrhea children with Streptococcus bovis as the dominant bacteria on admission disappeared with the improvement of their condition. We isolated and cultured the dominant flora of 16s rDNA, and determined the biochemistry reaction and the approximate full-length sequence of 16s rDNA. Two strains of Streptococcus Galulicus Pasteurus subspecies. We analyzed the whole genome sequence of streptococcus Paris isolated. It was found that there were 20 genomic islands in the genome of Streptococcus Paris, of which 7 (encoding glycosyltransferase) and 18 (encoding genes associated with fimbriae) were considered to be possible virulence islands, and 8 other possible pathogenicity genes were found. We report for the first time that Streptococcus Paris is the dominant bacteria in the intestinal flora of children with infectious diarrhea of unknown pathogens, and the proportion of Streptococcus Paris is decreasing with the course of treatment. Possible virulence islands and pathogenicity genes were identified. We isolated Streptococcus Bovis strains from feces of diarrhea patients and animal hosts and studied their molecular epidemiology.
【學(xué)位授予單位】：中國疾病預(yù)防控制中心
【學(xué)位級別】：博士
【學(xué)位授予年份】：2012
【分類號】：R725.1

【共引文獻】

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3 張曉燕;DnaK家族蛋白DnaJ介導(dǎo)豬鏈球菌2型的熱應(yīng)激及其對上皮細胞的黏附[D];浙江大學(xué);2014年

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1 王濤;豬鏈球菌穿血腦屏障毒力因子的篩選及鑒定[D];安徽醫(yī)科大學(xué);2013年

2 丁丹丹;豬鏈球菌2型cps2E基因缺失株構(gòu)建及生物學(xué)特性研究[D];華中農(nóng)業(yè)大學(xué);2014年

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本文編號：1509694