標題: 運用高通量定序分析陰道微生物菌相與陰道炎之相關性和精液菌相與不孕症之關聯性
Using high-throughput sequencing technology and bioinformatics analysis to discover the association between vaginal microbiota and bacterial vaginosis and the correlation between semen microbiota and infertility
作者: 翁順隆
Weng, Shun-Long
黃憲達
HUANG, HSIEN- DA
生物科技學系
關鍵字: 高通量定序;生物菌相;陰道炎;精液;不孕症;NGS;Microbiota;Semen;Vagina;Infertility
公開日期: 2015
摘要: 人體的各部位中存在著許多細菌,部分對人體有益,而部份則可能會傷害及影響人 體健康。目前已有研究發現,細菌不僅是造成陰道炎的主因,更與女性不孕症有關,此 外男性精菌症也與男性不孕症息息相關。根據衛生署的資料,台灣生育年齡的夫妻約有 15%有不孕的問題,不孕症儼然已成為一項嚴重的問題,而分析男女生殖道菌相與不孕 症之關聯性,將有助於找出不孕症之成因,提供未來診斷與治療的依據,因此是一相當 重要之研究。然而,過去細菌菌相(bacterial flora)之研究,大多使用培養(culture)或 PCR 方式進行分析,但上述方法只能針對數種細菌進行分析,且有許多細菌難以在人工培養 環境中存活,因此造成研究上之瓶頸。近年來隨著高通量定序(high‐throughput sequencing)技術快速發展,利用細菌 16S 宏觀基因體(Metagenomics)之分析方法提高 菌相證實的準確度,可大幅改善傳統菌相檢測之侷限性。 本研究將運用高通量定序技術,結合生物資訊分析,探討陰道菌相與細菌性陰道炎 的關係及精液菌相與男性不孕症之關聯性。本研究第一個主題是收集停經前與停經後婦 女之陰道檢體,運用高通量定序與生物資訊分析技術,建立台灣婦女陰道菌相資料庫, 並配合臨床特徵之診斷(Nugent Score、Amsel’s criteria 等),將樣本分為陰道炎與健康兩 類,藉此分析停經前與停經後婦女之陰道菌相組成與差異,並找出造成不同年齡層婦女 ii 之細菌性陰道炎的主要菌種與菌相特徵,研究結果發現細菌菌相與陰道檢體臨床特徵有 顯著之關聯性,研究成果不僅可作為細菌性陰道炎判斷與分類的依據,更可進一步作為 各年齡層細菌性陰道炎病患用藥之參考。 本研究第二個研究主題是運用高通量定序與生物資訊分析精液檢體,建立台灣男性 精液菌相資料庫,並配合臨床特徵之診斷(精蟲數量、活動力、型態與精蟲抗體等),將 樣本分為正常品質與低品質精液兩類,分析兩者菌相之差異,找出造成臨床特徵值差異 之菌種,研究結果顯示精液菌相可分為三種類型,且細菌菌相與精液品質有顯著之關聯 性,研究成果將有助於未來對男性不孕症的預防及治療。本研究成果可提供未來不孕症 夫妻檢測與治療的新方向。
Abstract The human body plays host to a vast array of bacteria which are harmful or beneficial. It has been know that vaginal microbiota is related to bacterial vaginosis and bateriospermia is correlated to male infertility. Most of the previous studies used culture method or PCR to analyze bacteria flora. Those methods could only focus on several bacteria. Moreover, most bacteria could not be cultured. Thus, it becomes a bottleneck of bacterial flora study. In recent years, limitation of traditional bacterial flora identification is improved as fast development of high‐throughput sequencing technology because of accuracy in identifying bacteria. In this study, we use high-throughput sequencing technology and bioinformatics analysis to investigate the relationship between vaginal bacterial flora and bacterial vaginosis and the correlations between semen bacterial flora and male infertility. The first aim of this study is to find the relationship between vaginal disorders and vaginal bacterial flora. The specimens were collected from pre‐menopause and menopause women. In order to find causes of vaginal disorders from different ages, we combine clinical iv examination criteria, such as Nugent Score and Amsel's criteria, to separate specimens into case and control group. The analysis results show that the specific patterns of vaginal flora are different in women from different age with bacterial vaginosis, and it could be used in prescription in bacterial vaginosis treatment. The secondary aim of this study is to analyze the relationship between semen flora, semen quality and male infertility. Here we present an analysis approach using next- generation sequencing technology and bioinformatics analysis to investigate the associations between bacterial communities and semen quality. The results suggest that the most abundant genera among all samples are Lactobacillus (19.9%), Pseudomonas (9.85%), Prevotella (8.51%) and Gardnerella (4.21%). The proportion of Lactobacillus and Gardnerella was significantly higher in the normal samples, while that of Prevotella was significantly higher in the low quality samples. The analysis results suggest seminal bacteria community types are highly associated with semen quality. The study significantly contributes to the current understanding of the etiology of male fertility.
URI: http://140.113.39.130/cdrfb3/record/nctu/#GT070187015
http://hdl.handle.net/11536/125776
Appears in Collections:Thesis