Dysbiosis in Inflammatory Bowel Disease

描述（由申请人提供）：炎症性肠病（IBD），即溃疡性结肠炎（DC）和克罗恩病（CD），是慢性、终身、复发性疾病，影响近100万美国人。尽管IBD中可能存在生态失调的许多线索，但IBD患者微生物区系的具体变化在很大程度上是未知的。扩增子长度异质性（ALH）是一种成熟的基于PCR的技术，可用作筛选工具来识别IBD患者细菌微生物群落的变化。因此，我们假设IBD患者的回结肠菌群与正常菌群相比，其微生物组成发生了改变。我们已经收集了初步数据，显示对照组和IBD之间以及IBD患者之间的统计学差异。因此，为了验证上述假设，我们提出了以下两个相互关联的科学目标：目的1。使用ALH鉴定与IBD相关的细菌指纹图谱。将在总共160名IBD患者和健康对照中确定ALH模式，并将使用多样性指数、多变量减少分析和聚类分析进行分析。将使用直方图、生态软件结合自定义PERL脚本以及监督和非监督自动模式识别系统来确定与IBD相关的ALH模式。目的2.利用分子克隆测序技术，确定脓毒症IBD相关ALH指纹图谱的细菌含量。通过采用多种生物信息学工具，将靶向样品的克隆和测序与细菌身份联系起来。意义该提案涉及首次使用复杂且高度可重复的分子生物学工具ALH来研究胃肠道中的微生物菌群。越来越多的人认识到微生物在健康和疾病中的重要性，包括IBD。使用来自环境微生物学的强大技术（如ALH）来表征人体微生物区系的研究可以在对胃肠道疾病的理解方面带来重大进展。ALH可能首次在医学上实现对微生物群落变化的实时调查，并可能提供将IBD与特定微生物模式联系起来的第一个证据。

DESCRIPTION (provided by applicant): Inflammatory Bowel Diseases (IBDs), namely ulcerative colitis (DC) and Crohn's disease (CD), are chronic, lifelong, relapsing illnesses, affecting close to 1 million Americans. Despite the many clues that a dysbiosis may exist in IBD, the specific changes in the microflora of IBD patients are largely unknown. Amplicon length heterogeneity (ALH) is a sophisticated and well established PCR based technology that can be used as a screening tool to identify changes in the Bacterial microflora of IBD patients. We therefore have hypothesized that the ileocolonic microflora in IBD has an altered microbial composition compared to normal microflora. We have gathered preliminary data that shows statistical differences between controls and IBD as well as within IBD patients. Therefore, to test the above hypothesis, we are proposing the following 2 inter-related scientific aims: AIM 1. Identify bacterial fingerprint patterns associated with IBD using ALH. ALH patterns will be determined in a total of 160 IBD patients and health controls and will be analyzed using diversity indeces, multivariate reduction analysis and cluster analysis. ALH patterns associated with IBD will be determined using histograms, ecological software in conjunction with custom PERL scripts as well as supervised and unsupervised automated pattern recognition systems. AIM 2.Determine the bacterial contents of putatively IBD associated ALH fingerprint patterns using molecular cloning and sequencing. Cloning and sequencing of targeted samples will be linked to bacterial identities by employing multiple bioinformatics tools. Significance. This proposal involves the first time use of a sophisticated and highly reproducible molecular biology tool, ALH, in the study of microflora in the Gl tract. There is a growing recognition of the importance of microflora in health and disease, including IBD. Studies that characterized microflora in human using powerful techniques from environmental microbiology such as ALH can bring about significant advances in the understanding of Gl tract illnesses. ALH may enable a real-time survey of microfloral changes for the first time in medicine and may provide the first evidence linking IBD with specific microbial patterns.