Molecular Analysis of Microbes in Chronic Bowel Disease

描述（由申请人提供）：本研究计划将 确定与炎症相关的组织的微生物成分 肠道疾病（IBD），在人类和动物系统，以确定 与疾病状态相关的微生物。人类炎症性肠道 疾病，溃疡性结肠炎（UC）和克罗恩病？的疾病（CD），是慢性的， 病因不明的毁灭性疾病有可能微生物， 也许是土著人，但迄今尚未发现， 过程尽管有证据表明鸟分枝杆菌亚种（Mycobacterium avium ssp.） 副结核杆菌（MAP）可能参与CD，微生物分析 由于临床上需要培养未知的微生物， 以便检测和识别它们。养殖技术往往 无效，通常低估了微生物的真实多样性， 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（例如宿主加相关的 微生物群）的聚合酶链反应，允许物种识别， 没有种植。我们建议分析微生物群落的 人IBD和正常胃肠道样品通过这种基于rRNA基因的 技术，以鉴定和表征IBD的候选微生物病原体。分子研究的结果将指导 直接尝试培养可疑的病原体，包括MAP，从患病的 组织中动物IBD模型的平行分析，包括牛约翰？S 疾病和IBD的啮齿动物模型，将进行，以验证 分子系统发育策略，提供深入了解微生物参与 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.