Division of Mucosal Immunology, Graduate School of Medicine, Shiga University of Medical Science, Seta Tsukinowa, Otsu 520-2192, Japan.
J Gastroenterol. 2011 Apr;46(4):479-86. doi: 10.1007/s00535-010-0368-4. Epub 2011 Jan 21.
Terminal restriction fragment length polymorphism (T-RFLP) analysis is a powerful tool to assess the diversity of a microbial community. In this study, we performed T-RFLP analysis of the fecal microbiota from patients with ulcerative colitis (UC) and those with Crohn's disease (CD).
Thirty-one patients with UC, 31 patients with CD, and 30 healthy individuals were enrolled. The polymerase chain reaction (PCR) products obtained from the 16S rRNA genes of fecal samples were digested with BslI, and T-RF lengths were determined.
The fecal microbial communities were classified into 5 clusters. Twenty-eight of the 30 healthy individuals and 17 of the 18 patients with inactive UC were classified into clusters I, II, and III, but these clusters included a small number of patients with active UC and inactive/active CD. In contrast, 8 of the 13 patients with active UC and the majority of CD patients (12 of the 16 patients with inactive CD, and 11 of the 15 patients with active CD) were included in clusters IV and V. Based on the BslI-digested T-RFLP database, the bacteria showed a significant decrease in the Clostridium family in patients with active UC and inactive/active CD. In contrast, Bacteroides were significantly increased in CD patients. No significant differences were observed between patients with active UC and those with active CD.
The fecal microbial communities of IBD patients were different from those of healthy individuals. The gut microbiota of patients with inactive UC tended to be closer to that of healthy individuals, suggesting different roles for the fecal microbiota in the pathophysiology of UC and CD.
末端限制性片段长度多态性(T-RFLP)分析是评估微生物群落多样性的有力工具。在这项研究中,我们对溃疡性结肠炎(UC)和克罗恩病(CD)患者的粪便微生物群进行了 T-RFLP 分析。
纳入 31 例 UC 患者、31 例 CD 患者和 30 名健康对照者。从粪便样本 16S rRNA 基因获得的聚合酶链反应(PCR)产物用 BslI 消化,并测定 T-RF 长度。
粪便微生物群落分为 5 个簇。30 名健康对照者中的 28 名和 18 名无活动 UC 患者中的 17 名被分为簇 I、II 和 III,但这些簇中包括少数活动 UC 患者和无活动/活动 CD 患者。相比之下,13 名活动 UC 患者中的 8 名和大多数 CD 患者(16 名无活动 CD 患者中的 12 名和 15 名活动 CD 患者中的 11 名)被归入簇 IV 和 V。基于 BslI 消化的 T-RFLP 数据库,活动 UC 和无活动/活动 CD 患者的梭菌属数量显著减少。相反,CD 患者的拟杆菌属显著增加。活动 UC 患者与活动 CD 患者之间未观察到显著差异。
IBD 患者的粪便微生物群落与健康对照者不同。无活动 UC 患者的肠道微生物群更倾向于健康对照者,提示粪便微生物群在 UC 和 CD 病理生理学中的作用不同。