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 in | Gut Vol. 55; no. 2; pp. 205 - 211 |
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Main Authors | , , , , , , , , , , , |
Format | Journal Article |
Language | English |
Published |
London
BMJ Publishing Group Ltd and British Society of Gastroenterology
01.02.2006
BMJ BMJ Publishing Group LTD BMJ Publishing Group BMJ Group |
Subjects | |
Online Access | Get full text |
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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. |
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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 |
Author_xml | – sequence: 1 givenname: C surname: Manichanh fullname: Manichanh, C organization: Department of Molecular Biology, IBMB-CSIC, Barcelona, Spain – sequence: 2 givenname: L surname: Rigottier-Gois fullname: Rigottier-Gois, L organization: Department of Molecular Biology, IBMB-CSIC, Barcelona, Spain – sequence: 3 givenname: E surname: Bonnaud fullname: Bonnaud, E organization: Department of Molecular Biology, IBMB-CSIC, Barcelona, Spain – sequence: 4 givenname: K surname: Gloux fullname: Gloux, K organization: Department of Molecular Biology, IBMB-CSIC, Barcelona, Spain – sequence: 5 givenname: E surname: Pelletier fullname: Pelletier, E organization: Department of Molecular Biology, IBMB-CSIC, Barcelona, Spain – sequence: 6 givenname: L surname: Frangeul fullname: Frangeul, L organization: Department of Molecular Biology, IBMB-CSIC, Barcelona, Spain – sequence: 7 givenname: R surname: Nalin fullname: Nalin, R organization: Department of Molecular Biology, IBMB-CSIC, Barcelona, Spain – sequence: 8 givenname: C surname: Jarrin fullname: Jarrin, C organization: Department of Molecular Biology, IBMB-CSIC, Barcelona, Spain – sequence: 9 givenname: P surname: Chardon fullname: Chardon, P organization: Department of Molecular Biology, IBMB-CSIC, Barcelona, Spain – sequence: 10 givenname: P surname: Marteau fullname: Marteau, P organization: Department of Molecular Biology, IBMB-CSIC, Barcelona, Spain – sequence: 11 givenname: J surname: Roca fullname: Roca, J organization: Department of Molecular Biology, IBMB-CSIC, Barcelona, Spain – sequence: 12 givenname: J surname: Dore fullname: Dore, J organization: Department of Molecular Biology, IBMB-CSIC, Barcelona, Spain |
BackLink | http://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=17402321$$DView record in Pascal Francis https://www.ncbi.nlm.nih.gov/pubmed/16188921$$D View this record in MEDLINE/PubMed https://hal.inrae.fr/hal-02667954$$DView record in HAL |
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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 |
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Notes | Correspondence to:
Dr J Dore
INRA-UEPSD, 78350 Jouy-en-Josas, France; joel.dore@jouy.inra.fr local:0550205 ark:/67375/NVC-M53D7CJV-1 href:gutjnl-55-205.pdf istex:DEEC691B1829E82F8F0E83A441AA12AD57FC78DA PMID:16188921 ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 14 content type line 23 |
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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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SubjectTerms | Adolescent Adult Bacteria Bacteria - classification Bacteria - genetics Bacteria - isolation & purification Bacterial Typing Techniques Biological and medical sciences Cloning CPL Crohn Disease - microbiology Crohn patient library Crohn's disease Deoxyribonucleic acid DNA DNA, Bacterial - genetics Ecosystem Ecosystems faecal microbiota Feces - microbiology Female Firmicutes FISH fluorescent in situ hybridisation Gastroenterology. Liver. Pancreas. Abdomen Gene Library Genes Genomes healthy subject library HSL Humans IBD In Situ Hybridization, Fluorescence Inflammatory Bowel Disease Life Sciences Male Medical sciences metagenome Microorganisms Molecular weight operational taxonomic unit Other diseases. Semiology OTU PCR Phylogenetics Phylogeny polymerase chain reaction RNA, Bacterial - genetics RNA, Ribosomal, 16S - genetics saline sodium citrate SDS sodium dodecyl sulphate SSC Stomach. Duodenum. Small intestine. Colon. Rectum. Anus Studies |
Title | Reduced diversity of faecal microbiota in Crohn’s disease revealed by a metagenomic approach |
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