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

BackgroundTerminal restriction fragment length polymorphism (T-RFLP) analysis is a powerful tool 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 ulce...

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Vydáno v:Inflammatory bowel diseases Ročník 13; číslo 8; s. 955 - 962
Hlavní autoři: Andoh, Akira, Sakata, Shinji, Koizumi, Yuhsuke, Mitsuyama, Keiichi, Fujiyama, Yoshihide, Benno, Yoshimi
Médium: Journal Article
Jazyk:angličtina
Vydáno: Oxford, UK Oxford University Press 01.08.2007
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Témata:
Adult
Age
Bacteria
Bacteroides
Colitis, Ulcerative - microbiology
commensals
DNA, Bacterial - analysis
Enzymes
Eubacterium
Feces
Feces - microbiology
Female
Fusobacterium
Humans
IBD
Inflammatory bowel disease
Inflammatory bowel diseases
intestinal microbiota
Intestine
Lactobacillus
Male
Microbiota
Polymerase Chain Reaction
Polymorphism
Polymorphism, Restriction Fragment Length
pouchitis
Proctitis
Remission
Restriction fragment length polymorphism
rRNA 16S
Ruminococcus
Ulcerative colitis
intestinal microbiota
IBD
commensals
pouchitis
ISSN:1078-0998, 1536-4844
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Shrnutí:BackgroundTerminal restriction fragment length polymorphism (T-RFLP) analysis is a powerful tool 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).MethodsForty-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.ResultsThe fecal microbial communities were classified in 8 clusters. Almost all the healthy individuals (39 of 46) were included in cluster I, 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.ConclusionsT-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.
Bibliografie:Akira Andoh and Shinji Sakata contributed equally to this work.
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ISSN:1078-0998
1536-4844
DOI:10.1002/ibd.20151