運用宏觀基因體分析平台建立人體菌相圖譜及細菌與疾病之風險評估模型

Abstract

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。近年來，高通量次世代定序技術已能準確的應用於微生物及細菌的鑑定。此研究致力於發展一個運用高通量16S rRNA定序技術建立人體菌相圖譜及建構細菌相關疾病之風險評估模型之分析平台。藉由文獻及公開的序列資料庫蒐集細菌之16S rRNA序列建構細菌16s rRNA序列資料庫，並搭配序列品質控管、奇美拉序列偵測、序列分類及序列比對來對於人體中的菌種鑑定。目前此研究蒐集了174台灣健康人類糞便樣本及針對七項腸胃道疾病找出78個疾病相關之細菌特徵，建構細菌相關疾病之風險評估模型評估腸胃道相關疾病之風險。此研究亦於不同時期蒐集了81個台灣人糞便樣本及51個台灣人糞便樣本用來初步測試此平台的應用能力。目前鎖定腸胃道相關疾病、陰道相關疾病及不孕症蒐集相關細菌特徵，並搜集糞便、陰道及精液樣本進行case-control研究，以期能夠找到新的疾病相關之細菌特徵，進而延伸細菌相關疾病之風險評估模型。此平台能夠大量分析人體部位之細菌族群，並藉由蒐集健康台灣人樣本建立台灣人菌相圖譜，進而利用細菌相關疾病之風險評估模型分析應用於臨床檢測及研究，了解那些細菌與人體健康息息相關並藉由菌相的了解發展新的治療方法。

The human body plays host to a vast array of bacteria which are harmful or beneficial. Next generation sequencing technology have increased its accuracy in identifying bacteria. This work develops a novel platform for Metagenomics analysis, bacterial disease risk evaluation model construction and microbiome characterization in human using high-throughput 16S rRNA sequencing. A database that recorded the bacteria 16s rRNA sequences from literature and public database was constructed, along with quality filtering, chimeric sequence detection, sequence clustering and assigning the taxonomy of the sequenced 16S rRNA sequences. A bacteria disease risk model for seven diseases was constructed based on 174 samples and 78 bacterial disease risk biomarkers. Applicability of the proposed platform is demonstrated by collecting the microbiome in human gut of 81 samples and 53 samples with BMI. For analyzing bacteria related disease, this work expects to collect more bacterial disease biomarkers from human body. Additionally, we design case-control study to finding novel association between bacteria and disease for extending bacterial disease risk evaluation model. The proposed platform provides a relatively easy means of identifying a certain amount of bacteria and evaluating bacterial disease risk based on Taiwanese control group for clinical microbiology applications. Detecting how bacteria inhabit humans and affect their health significantly contributes to develop a diagnosis and treatment method.