Molecular Analysis of Microbes in Chronic Bowel Disease

描述(由申请者提供)：这项拟议的研究计划将 测定炎症相关组织的微生物成分 人类和动物系统中的肠道疾病(IBD)，以便识别 与疾病状态相关的微生物。人类炎症性肠病 溃疡性结肠炎(UC)和克罗恩-S病(CD)是慢性疾病， 原因不明的毁灭性疾病。有可能微生物， 可能是土著的，而且到目前为止还没有发现，参与了这种疾病 进程。尽管有证据表明，禽类分枝杆菌SSP. 微生物学分析表明，副结核(MAP)可能与CD有关 受到培养未知生物的临床需求的严重影响 以便检测和识别它们。养殖技术经常被 效率低下，而且通常低估了细菌的真实多样性 天然样品。核糖体RNA(RRNA)基因的系统发育分析， 从混合群落基因组DNA扩增(例如，寄主+相关 微生物群)，通过聚合酶链式反应，允许在 缺乏修养。我们建议分析以下微生物群落： 以此rRNA基因为基础的人IBD和正常胃肠道标本 技术，以确定和表征IBD的候选微生物病原体。分子研究的结果将指导 定向尝试培养疑似病原体，包括MAP 纸巾。牛传染性法氏囊病动物模型的平行分析 将进行IBD的疾病和啮齿动物模型，以验证 分子系统发育策略，提供了对微生物参与 IBD的发病机制，并指导选择合适的组织进行分析 在人类IBD中。

DESCRIPTION (provided by applicant): This proposed research program will determine the microbial constituents of tissues associated with inflammatory bowel diseases (IBD), in both human and animal systems, in order to identify microbes associated with the disease-states. The human inflammatory bowel diseases, ulcerative colitis (UC) and Crohn?s disease (CD), are chronic, devastating diseases of unknown etiology. It is possible that microorganisms, perhaps indigenous and so far undiscovered, participate in the disease process. Although some evidence suggests that Mycobacterium avium ssp. paratuberculosis (MAP) may be involved in CD, microbiological analyses have been critically compromised by the clinical need to culture unknown organisms in order to detect and identify them. Culture techniques are frequently ineffective and usually underestimate the true diversity of microbes in natural samples. The phylogenetic analysis of ribosomal RNA (rRNA) genes, amplified from mixed community genomic DNA (e.g. host plus associated microbiota) by polymerase chain reaction, allows species identification in the absence of cultivation. We propose to analyze the microbial communities of both human IBD and normal gastrointestinal samples by this rRNA gene-based technology in order to identify and characterize candidate microbial etiological agents of IBD. The results of molecular studies will guide directed attempts to culture suspected pathogens, including MAP, from diseased tissues. Parallel analyses of animal IBD models, including bovine Johne?s disease and rodent models of IBD, will be conducted in order to validate the molecular-phylogenetic strategy, provide insight into microbial involvement in IBD pathogenesis, and guide the choice of appropriate tissues to be analyzed in human IBD.