Reduced diversity of faecal microbiota in Crohn’s disease revealed by a metagenomic approach

Background and aim: A role for the intestinal microbial community (microbiota) in the onset and chronicity of Crohn’s disease (CD) is strongly suspected. However, investigation of such a complex ecosystem is difficult, even with culture independent molecular approaches. Methods: We used, for the fir...

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Published inGut Vol. 55; no. 2; pp. 205 - 211
Main Authors Manichanh, C, Rigottier-Gois, L, Bonnaud, E, Gloux, K, Pelletier, E, Frangeul, L, Nalin, R, Jarrin, C, Chardon, P, Marteau, P, Roca, J, Dore, J
Format Journal Article
LanguageEnglish
Published London BMJ Publishing Group Ltd and British Society of Gastroenterology 01.02.2006
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Abstract Background and aim: A role for the intestinal microbial community (microbiota) in the onset and chronicity of Crohn’s disease (CD) is strongly suspected. However, investigation of such a complex ecosystem is difficult, even with culture independent molecular approaches. Methods: We used, for the first time, a comprehensive metagenomic approach to investigate the full range of intestinal microbial diversity. We used a fosmid vector to construct two libraries of genomic DNA isolated directly from faecal samples of six healthy donors and six patients with CD. Bacterial diversity was analysed by screening the two DNA libraries, each composed of 25 000 clones, for the 16S rRNA gene by DNA hybridisation. Results: Among 1190 selected clones, we identified 125 non-redundant ribotypes mainly represented by the phyla Bacteroidetes and Firmicutes. Among the Firmicutes, 43 distinct ribotypes were identified in the healthy microbiota, compared with only 13 in CD (p<0.025). Fluorescent in situ hybridisation directly targeting 16S rRNA in faecal samples analysed individually (n = 12) confirmed the significant reduction in the proportion of bacteria belonging to this phylum in CD patients (p<0.02). Conclusion: The metagenomic approach allowed us to detect a reduced complexity of the bacterial phylum Firmicutes as a signature of the faecal microbiota in patients with CD. It also indicated the presence of new bacterial species.
AbstractList Background and aim: A role for the intestinal microbial community (microbiota) in the onset and chronicity of Crohn’s disease (CD) is strongly suspected. However, investigation of such a complex ecosystem is difficult, even with culture independent molecular approaches. Methods: We used, for the first time, a comprehensive metagenomic approach to investigate the full range of intestinal microbial diversity. We used a fosmid vector to construct two libraries of genomic DNA isolated directly from faecal samples of six healthy donors and six patients with CD. Bacterial diversity was analysed by screening the two DNA libraries, each composed of 25 000 clones, for the 16S rRNA gene by DNA hybridisation. Results: Among 1190 selected clones, we identified 125 non-redundant ribotypes mainly represented by the phyla Bacteroidetes and Firmicutes. Among the Firmicutes, 43 distinct ribotypes were identified in the healthy microbiota, compared with only 13 in CD (p<0.025). Fluorescent in situ hybridisation directly targeting 16S rRNA in faecal samples analysed individually (n = 12) confirmed the significant reduction in the proportion of bacteria belonging to this phylum in CD patients (p<0.02). Conclusion: The metagenomic approach allowed us to detect a reduced complexity of the bacterial phylum Firmicutes as a signature of the faecal microbiota in patients with CD. It also indicated the presence of new bacterial species.
A role for the intestinal microbial community (microbiota) in the onset and chronicity of Crohn's disease (CD) is strongly suspected. However, investigation of such a complex ecosystem is difficult, even with culture independent molecular approaches. We used, for the first time, a comprehensive metagenomic approach to investigate the full range of intestinal microbial diversity. We used a fosmid vector to construct two libraries of genomic DNA isolated directly from faecal samples of six healthy donors and six patients with CD. Bacterial diversity was analysed by screening the two DNA libraries, each composed of 25,000 clones, for the 16S rRNA gene by DNA hybridisation. Among 1190 selected clones, we identified 125 non-redundant ribotypes mainly represented by the phyla Bacteroidetes and Firmicutes. Among the Firmicutes, 43 distinct ribotypes were identified in the healthy microbiota, compared with only 13 in CD (p<0.025). Fluorescent in situ hybridisation directly targeting 16S rRNA in faecal samples analysed individually (n=12) confirmed the significant reduction in the proportion of bacteria belonging to this phylum in CD patients (p<0.02). The metagenomic approach allowed us to detect a reduced complexity of the bacterial phylum Firmicutes as a signature of the faecal microbiota in patients with CD. It also indicated the presence of new bacterial species.
A role for the intestinal microbial community (microbiota) in the onset and chronicity of Crohn's disease (CD) is strongly suspected. However, investigation of such a complex ecosystem is difficult, even with culture independent molecular approaches.BACKGROUND AND AIMA role for the intestinal microbial community (microbiota) in the onset and chronicity of Crohn's disease (CD) is strongly suspected. However, investigation of such a complex ecosystem is difficult, even with culture independent molecular approaches.We used, for the first time, a comprehensive metagenomic approach to investigate the full range of intestinal microbial diversity. We used a fosmid vector to construct two libraries of genomic DNA isolated directly from faecal samples of six healthy donors and six patients with CD. Bacterial diversity was analysed by screening the two DNA libraries, each composed of 25,000 clones, for the 16S rRNA gene by DNA hybridisation.METHODSWe used, for the first time, a comprehensive metagenomic approach to investigate the full range of intestinal microbial diversity. We used a fosmid vector to construct two libraries of genomic DNA isolated directly from faecal samples of six healthy donors and six patients with CD. Bacterial diversity was analysed by screening the two DNA libraries, each composed of 25,000 clones, for the 16S rRNA gene by DNA hybridisation.Among 1190 selected clones, we identified 125 non-redundant ribotypes mainly represented by the phyla Bacteroidetes and Firmicutes. Among the Firmicutes, 43 distinct ribotypes were identified in the healthy microbiota, compared with only 13 in CD (p<0.025). Fluorescent in situ hybridisation directly targeting 16S rRNA in faecal samples analysed individually (n=12) confirmed the significant reduction in the proportion of bacteria belonging to this phylum in CD patients (p<0.02).RESULTSAmong 1190 selected clones, we identified 125 non-redundant ribotypes mainly represented by the phyla Bacteroidetes and Firmicutes. Among the Firmicutes, 43 distinct ribotypes were identified in the healthy microbiota, compared with only 13 in CD (p<0.025). Fluorescent in situ hybridisation directly targeting 16S rRNA in faecal samples analysed individually (n=12) confirmed the significant reduction in the proportion of bacteria belonging to this phylum in CD patients (p<0.02).The metagenomic approach allowed us to detect a reduced complexity of the bacterial phylum Firmicutes as a signature of the faecal microbiota in patients with CD. It also indicated the presence of new bacterial species.CONCLUSIONThe metagenomic approach allowed us to detect a reduced complexity of the bacterial phylum Firmicutes as a signature of the faecal microbiota in patients with CD. It also indicated the presence of new bacterial species.
Author Bonnaud, E
Frangeul, L
Jarrin, C
Rigottier-Gois, L
Roca, J
Gloux, K
Pelletier, E
Nalin, R
Marteau, P
Chardon, P
Dore, J
Manichanh, C
AuthorAffiliation R Nalin , C Jarrin , LibraGen, Bat Canal Biotech I, Toulouse, France
P Marteau , Gastroenterology Department, European Hospital Georges Pompidou, Paris, France
L Frangeul , Genopole, Institut Pasteur, Paris, France
C Manichanh , L Rigottier‐Gois , E Bonnaud , K Gloux , J Dore , Unité d'Ecologie et de Physiologie du système Digestif, INRA Jouy‐en‐Josas, France
P Chardon , UMR INRA CEA radiobiologie et étude du génome, INRA Domaine de Vilvert, Jouy‐en‐Josas, Cedex France
J Roca , Department of Molecular Biology, IBMB‐CSIC, Barcelona, Spain
E Pelletier , Genoscope, Centre National de Séquençage, Evry cedex, France
AuthorAffiliation_xml – name: P Chardon , UMR INRA CEA radiobiologie et étude du génome, INRA Domaine de Vilvert, Jouy‐en‐Josas, Cedex France
– name: E Pelletier , Genoscope, Centre National de Séquençage, Evry cedex, France
– name: L Frangeul , Genopole, Institut Pasteur, Paris, France
– name: P Marteau , Gastroenterology Department, European Hospital Georges Pompidou, Paris, France
– name: R Nalin , C Jarrin , LibraGen, Bat Canal Biotech I, Toulouse, France
– name: C Manichanh , L Rigottier‐Gois , E Bonnaud , K Gloux , J Dore , Unité d'Ecologie et de Physiologie du système Digestif, INRA Jouy‐en‐Josas, France
– name: J Roca , Department of Molecular Biology, IBMB‐CSIC, Barcelona, Spain
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IsDoiOpenAccess true
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Issue 2
Keywords Crohn disease
Gastroenterology
Digestive diseases
Intestinal disease
Surgical approach
Feces
Inflammatory disease
PHYLOGENY
16S rRNA
CROHN'S DISEASE
FAECAL MICROBIOTA
METAGENOME
FIRMICUTES
FLUORESCENT IN SITU HIBRIDIZATION
Language English
License CC BY 4.0
Distributed under a Creative Commons Attribution 4.0 International License: http://creativecommons.org/licenses/by/4.0
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Notes Correspondence to:
 Dr J Dore
 INRA-UEPSD, 78350 Jouy-en-Josas, France; joel.dore@jouy.inra.fr
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SSID ssj0008891
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Snippet Background and aim: A role for the intestinal microbial community (microbiota) in the onset and chronicity of Crohn’s disease (CD) is strongly suspected....
A role for the intestinal microbial community (microbiota) in the onset and chronicity of Crohn's disease (CD) is strongly suspected. However, investigation of...
Background and aim: A role for the intestinal microbial community (microbiota) in the onset and chronicity of Crohn's disease (CD) is strongly suspected....
BACKGROUND: and aim: A role for the intestinal microbial community (microbiota) in the onset and chronicity of Crohn's disease (CD) is strongly suspected....
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StartPage 205
SubjectTerms Adolescent
Adult
Bacteria
Bacteria - classification
Bacteria - genetics
Bacteria - isolation & purification
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Title Reduced diversity of faecal microbiota in Crohn’s disease revealed by a metagenomic approach
URI http://gut.bmj.com/content/55/2/205.full
https://api.istex.fr/ark:/67375/NVC-M53D7CJV-1/fulltext.pdf
https://www.ncbi.nlm.nih.gov/pubmed/16188921
https://www.proquest.com/docview/1779335330
https://www.proquest.com/docview/19433594
https://www.proquest.com/docview/70682916
https://hal.inrae.fr/hal-02667954
https://pubmed.ncbi.nlm.nih.gov/PMC1856500
Volume 55
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