Terminal restriction fragment length polymorphism analysis of the diversity of fecal microbiota in patients with ulcerative colitis

被引:90
作者
Andoh, Akira
Sakata, Shinji
Koizumi, Yuhsuke
Mitsuyama, Keiichi
Fujiyoma, Yoshihide
Benno, Yoshimi
机构
[1] Shiga Univ Med Sci, Dept Internal Med, Otsu, Shiga 5202191, Japan
[2] RIKEN, Bioresource Ctr, Microbe Div Japan Collect Microorganisms, Wako, Saitama 35101, Japan
[3] Kurume Univ, Sch Med, Dept Med, Kurume, Fukuoka 830, Japan
关键词
commensals; IBD; intestinal microbiota; pouchitis;
D O I
10.1002/ibd.20151
中图分类号
R57 [消化系及腹部疾病];
学科分类号
摘要
Background: Terminal restriction fragment length polymorphism (T-RFLP) analysis is a powerful toot to assess the diversity of complexed microbiota. This permits rapid comparison of microbiota from many samples. In this study, we performed T-RFLP analysis of the fecal microbiota from patients with ulcerative colitis (UC). Conclusions: T-RFLP analyses showed that the diversity of fecal microbiota in patients with UC was different from that in healthy individuals. Unclassified bacteria, as well as known bacteria, can contribute to alterations in the bacterial diversity of UC patients. Methods: Forty-four patients with UC (23 women and 21 men, median age 25 years) and 46 healthy individuals (25 women and 21 men, median age 34 years) were enrolled in this study. DNA was extracted from their stool samples, and the 16S rRNA genes were amplified by PCR. The PCR products were then digested with HhaI and/or MspI restriction enzymes, and the length of the T-RF was determined. Results: The fecal microbial communities were classified in 8 clusters. Almost all the healthy individuals (39 of 46) were included in cluster 1, and most of the UC patients could be divided into the other 7 clusters, indicating that fecal bacterial communities are different between healthy individuals and active UC patients. Some T-RFs, derived from the unclassified bacteria, Ruminococcus, Eubacterium, Fusobacterium, gammaproteobacteria, unclassified Bacteroides, and unclassified Lactobacillus, were detected in the UC patients, but not in the healthy individuals. The T-RFLP patterns were also different between the active patients and inactive (remission) patients. The T-RF derived from the unclassified bacteria, Ruminococcus and Eubacterium, and the T-RFs derived from the unclassified bacteria, Eubacterium, and Fusobacterium were predominantly detected in the active patients not the inactive patients. In contrast, the T-RFs derived from Lactobacillus and unclassified Lactobacillus were more predominant in the inactive (remission) patients. In 4 patients with proctitis, the pattern of fecal microbial diversity was very similar. Conclusions: T-RFLP analyses showed that the diversity of fecal microbiota in patients with UC was different from that in healthy individuals. Unclassified bacteria, as well as known bacteria, can contribute to alterations in the bacterial diversity of UC patients.
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页码:955 / 962
页数:8
相关论文
共 38 条
  • [1] The bacteriology of biopsies differs between newly diagnosed, untreated, Crohn's disease and ulcerative colitis patients
    Bibiloni, Rodrigo
    Mangold, Marco
    Madsen, Karen L.
    Fedorak, Richard N.
    Tannock, Gerald W.
    [J]. JOURNAL OF MEDICAL MICROBIOLOGY, 2006, 55 (08) : 1141 - 1149
  • [2] Gut-associated bacterial microbiota in paediatric patients with inflammatory bowel disease
    Conte, M. P.
    Schippa, S.
    Zamboni, I.
    Penta, M.
    Chiarini, F.
    Seganti, L.
    Osborn, J.
    Falconieri, P.
    Borrelli, O.
    Cucchiara, S.
    [J]. GUT, 2006, 55 (12) : 1760 - 1767
  • [3] Franks AH, 1998, APPL ENVIRON MICROB, V64, P3336
  • [4] ROLE OF THE FECAL STREAM IN THE MAINTENANCE OF CROHNS COLITIS
    HARPER, PH
    LEE, ECG
    KETTLEWELL, MGW
    BENNETT, MK
    JEWELL, DP
    [J]. GUT, 1985, 26 (03) : 279 - 284
  • [5] Diversity of the Clostridium coccoides group in human fecal microbiota as determined by 16S rRNA gene library
    Hayashi, H
    Sakamoto, M
    Kitahara, M
    Benno, Y
    [J]. FEMS MICROBIOLOGY LETTERS, 2006, 257 (02) : 202 - 207
  • [6] Molecular analysis of jejunal, ileal, caecal and recto-sigmoidal human colonic microbiota using 16S rRNA gene libraries and terminal restriction fragment length polymorphism
    Hayashi, H
    Takahashi, R
    Nishi, T
    Sakamoto, M
    Benno, Y
    [J]. JOURNAL OF MEDICAL MICROBIOLOGY, 2005, 54 (11) : 1093 - 1101
  • [7] Molecular analysis of fecal microbiota in elderly individuals using 16S rDNA library and T-RFLP
    Hayashi, H
    Sakamoto, M
    Kitahara, M
    Benno, Y
    [J]. MICROBIOLOGY AND IMMUNOLOGY, 2003, 47 (08) : 557 - 570
  • [8] Phylogenetic analysis of the human gut microbiota using 16S rDNA clone libraries and strictly anaerobic culture-based methods
    Hayashi, H
    Sakamoto, M
    Benno, Y
    [J]. MICROBIOLOGY AND IMMUNOLOGY, 2002, 46 (08) : 535 - 548
  • [9] Alteration of Vβ usage and cytokine production of CD4+ TCR ββ homodimer T cells by elimination of Bacteroides vulgatus prevents colitis in TCR α-chain-deficient mice
    Kishi, D
    Takahashi, I
    Kai, Y
    Tamagawa, H
    Iijima, H
    Obunai, S
    Nezu, R
    Ito, T
    Matsuda, H
    Kiyono, H
    [J]. JOURNAL OF IMMUNOLOGY, 2000, 165 (10) : 5891 - 5899
  • [10] Kitts C L, 2001, Curr Issues Intest Microbiol, V2, P17