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系統識別號 U0026-0410201214453000
論文名稱(中文) 大腸桿菌的可能醣類代謝基因之流行病學分析
論文名稱(英文) Epidemiological analysis of potential sugar-metabolic genes in Escherichia coli
校院名稱 成功大學
系所名稱(中) 分子醫學研究所
系所名稱(英) Institute of Molecular Medicine
學年度 101
學期 1
出版年 101
研究生(中文) 李馨
研究生(英文) Hsin Li
學號 t16991038
學位類別 碩士
語文別 中文
論文頁數 60頁
口試委員 指導教授-鄧景浩
口試委員-王憲威
口試委員-朱紀實
中文關鍵字 泌尿道致病性大腸桿菌  泌尿道感染  毒力因子  phylogenetic group  醣類代謝基因 
英文關鍵字 Uropathogenic Escherichia coli (UPEC)  Urinary tract infection (UTI)  virulence genes  phylogenetic groups  sugar metabolic genes 
學科別分類
中文摘要 泌尿道致病性大腸桿菌是泌尿道感染中最常見的病原菌之一。藉由流行病學的研究,我們可以找出其分佈和泌尿道感染具有相關性的大腸桿菌基因。這些與泌尿道感染相關的細菌基因可能是毒力因子,或是和毒力因子有genetic linkage的基因。因此,有這類特性的基因將有潛力成為區別泌尿道致病性大腸桿菌和非致病性大腸桿菌的標誌,或可能當作用來發展預防或治療泌尿道感染新策略的標的基因。
在本篇研究裡,我以流行病學的角度去探討五個大腸桿菌的可能醣類代謝基因 (c3408、c4487、c4758、c4496、c4984)和泌尿道感染、phylogenetic groups、抗藥性、以及彼此之間和已知毒力因子間分佈的相關性。結果顯示,c3408、c4487、c4758、c4984和泌尿道感染有顯著的相關性,但和膽道感染引起菌血症無顯著相關,因此這些基因可能和腸道外感染具疾病專一性。此外,這些可能醣類代謝基因在phylogenetic groups的分佈以及和抗藥性的相關性,和大部分的已知的泌尿道相關毒力因子一樣。它們大部分都集中在phylogenetic group B2,次之為group D; 且和Quinolones及Fluoroquinolones類抗生素抗藥性的關係呈現負相關性。而c3408、c4487、c4758、c4984除了和彼此之間呈現分佈上的正相關性外,也和大部分的已知毒力因子呈現正相關; 雖然c4496只和c4487呈現分佈上的負相關,而和其他可能醣類代謝基因無顯著相關性,但它和已知毒力因子的sisB、afa/draBC呈現正相關。而根據前人研究結果,c3408已經在動物感染實驗中被發現和泌尿道感染致病機轉相關,c4487和c4496在禽致病性大腸桿菌中的homolog也已經在雞感染實驗中被證明和致病相關。綜合我們流行病學的結果以及前人相關動物實驗,c3408、c4487、c4758、c4984將有潛力變成鑑定、預防及治療大腸桿菌泌尿道感染的標的基因。
另外,我也探討了在人類及牛大腸桿菌中可能醣類代謝基因及已知毒力因子的盛行率、phylogenetic groups及抗藥性的差異。結果顯示這兩類大腸桿菌群的phylogenetic groups組成比例不同,可能是造成大部分的可能醣類代謝及已知毒力因子在人類分離大腸桿菌分佈比例顯著高於牛的原因。c4758和已知毒力因子eco274、traT、ompT在牛乳腺炎致病性大腸桿菌中的比例明顯較高於其他基因,這些基因可能對於牛致病扮演角色。而在抗藥性部分,我發現Nalidixic acid抗藥性菌株比例在人類顯著高於牛分離大腸桿菌。這個結果可以提供對於抗生素使用上的建議。
英文摘要 Uropathogenic Escherichia coli (UPEC) is one of the most common pathogens of urinary tract infections (UTIs). Novel UTI-associated genes may be identified through epidemiological investigation. E. coli genes epidemiologically associated with UTIs may be the potential virulence genes itself or has a genetic linkage to such a gene. Therefore, UTI-associated E. coli genes are potential targets for developing novel strategies to treat and/or prevent E. coli-caused UTI, and being the marker to differentiate UPEC from non-UPEC isolates.
In this study, I evaluated the epidemiological association of five E. coli potential sugar-metabolic genes’ distributions (PSMGs: c3408、c4487、c4758、c4496、c4984) with UTI、 bacterial phylogenetic groups、antibiotic resistance, and known UTI-associated virulence genes. I found that the distributions of c3408、c4487、c4758、c4984 were significantly associated with UTI, but not with biliary tract infection. This result suggests that the associations of these PSMGs with E. coli-caused extraintestinal diseases may be syndrome-dependent. The patterns of these PSMGs' phylogenetic distributions and their associations with antibiotic resistance were similar to those of the majority of the known virulence genes. PSMGs mainly belong to phylogenetic groups B2, and, to a lesser extent, D; PSMGs exhibited negative association with Quinolones and Fluoroquinolones resistance. Besides, c3408、c4487、c4758 and c4984 showed positive associations with each other as well as most of the known virulence genes. c4496 was negatively associated with c4487 and positively associated with two of the known virulence genes, sisB and afa/draBC. c3408 has been shown to be involved in the pathogenesis of UPEC in the murine model of UTI. Also, the homolog gene of c4487 in avian pathogenic E. coli (APEC) has been shown to be involved in pathogenesis in chicken infection. According to our and others' studies, c3408、c4487、c4758、and c4984 are the potential target genes to develop diagnostic, preventive and/or therapeutic strategies of E. coli-caused UTI.
In addition, the distributions of PSMGs, known virulence genes, bacterial phylogenetic groups and antibiotic resistance were compared between human UPEC and bovine mastitis-associated E. coli (BMEC) strains. I found that the prevalence of most PSMGs and known virulence genes in human E. coli isolates were significant higher than those in BMEC. This difference may be due to the distinct combinations of the phylogenetic groups between these two types of pathogenic E. coli isolates. There are some genes’ prevalence in BMEC are higher than the others, such as c4758, eco274, traT and ompT. These genes may also play a role in pathogenesis of BMEC. In addition, I found that the distribution of the Nalidixic acid resistant strains in human UPEC was significantly higher than that in BMEC. This finding may provide a hint for antibiotic usage.
論文目次 中文摘要I
英文摘要III
誌謝 V
目錄VI
表目錄VIII
縮寫表IX
第一章、 緒論1
第一節 大腸桿菌 1
第二節 泌尿道感染 3
第三節 乳腺炎 6
第四節 腸桿菌科的醣類運輸系統和致病的相關性 6
第五節 實驗目的 7
第二章、 材料與方法 8
第一節 大腸桿菌之來源、培養、保存 8
第二節 聚合酶連鎖反應 8
第三節 洋菜膠電泳 9
第四節 藥敏性實驗 10
第五節 統計方法 11
第三章、 實驗結果12
第一節 UPEC 的可能醣類代謝基因與UTI 的相關性 12
第二節 PSMGs 與phylogenetic groups 的相關性 13
第三節 PSMGs 和彼此之間以及已知毒力因子分佈的相關性 15
第四節 UPEC 的PSMGs 與抗藥性的相關性 15
第五節 PSMGs 和宿主種類的相關性 17
第四章、 結果討論21
第五章、 參考文獻31
表43
附錄60
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