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Molecular analysis of faecal and duodenal samples reveals significantly higher prevalence and numbers of Pseudomonas aeruginosa in irritable bowel syndrome.

Abstract

Intestinal microbiota may play a role in the pathophysiology of irritable bowel syndrome (IBS). In this case-control study, mucosa-associated small intestinal and faecal microbiota of IBS patients and healthy subjects were analysed using molecular-based methods. Duodenal mucosal brush and faecal samples were collected from 37 IBS patients and 20 healthy subjects. The bacterial 16S rRNA gene was amplified and analysed using PCR denaturing gradient gel electrophoresis (DGGE). Pooled average DGGE profiles of all IBS patients and all healthy subjects from both sampling sites were generated and fingerprints of both groups were compared. The DGGE band fragments which were confined to one group were further characterized by sequence analysis. Quantitative real-time PCR (q-PCR) was used to quantify the disease-associated microbiota. Averaged DGGE profiles of both groups were identical for 78.2 % in the small intestinal samples and for 86.25 % in the faecal samples. Cloning and sequencing of the specific bands isolated from small intestinal and faecal DGGE patterns of IBS patients showed that 45.8 % of the clones belonged to the genus Pseudomonas, of which Pseudomonas aeruginosa was the predominant species. q-PCR analysis revealed higher levels (P<0.001) of P. aeruginosa in the small intestine of IBS patients (8.3 %±0.950) than in the small intestine of healthy subjects (0.1 %±0.069). P. aeruginosa was also significantly (P<0.001) more abundant (2.34 %±0.31) in faeces of IBS patients than in faeces of healthy subjects (0.003 %±0.0027). This study shows that P. aeruginosa is detected more frequently and at higher levels in IBS patients than in healthy subjects, suggesting its potential role in the pathophysiology of IBS.

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  • Authors+Show Affiliations

    ,

    Gastrointestinal Research Unit, Departments of Gastroenterology and Surgery, University Medical Center Utrecht, Utrecht, The Netherlands. AngeleKerckhoffs@hotmail.com

    , , , , ,

    Source

    Journal of medical microbiology 60:Pt 2 2011 Feb pg 236-45

    MeSH

    Adult
    Duodenum
    Electrophoresis, Polyacrylamide Gel
    Feces
    Female
    Humans
    Irritable Bowel Syndrome
    Male
    Molecular Typing
    Nucleic Acid Denaturation
    Polymerase Chain Reaction
    Prevalence
    Pseudomonas aeruginosa
    RNA, Bacterial
    RNA, Ribosomal, 16S

    Pub Type(s)

    Journal Article

    Language

    eng

    PubMed ID

    20947663

    Citation

    Kerckhoffs, Angèle P M., et al. "Molecular Analysis of Faecal and Duodenal Samples Reveals Significantly Higher Prevalence and Numbers of Pseudomonas Aeruginosa in Irritable Bowel Syndrome." Journal of Medical Microbiology, vol. 60, no. Pt 2, 2011, pp. 236-45.
    Kerckhoffs AP, Ben-Amor K, Samsom M, et al. Molecular analysis of faecal and duodenal samples reveals significantly higher prevalence and numbers of Pseudomonas aeruginosa in irritable bowel syndrome. J Med Microbiol. 2011;60(Pt 2):236-45.
    Kerckhoffs, A. P., Ben-Amor, K., Samsom, M., van der Rest, M. E., de Vogel, J., Knol, J., & Akkermans, L. M. (2011). Molecular analysis of faecal and duodenal samples reveals significantly higher prevalence and numbers of Pseudomonas aeruginosa in irritable bowel syndrome. Journal of Medical Microbiology, 60(Pt 2), pp. 236-45. doi:10.1099/jmm.0.022848-0.
    Kerckhoffs AP, et al. Molecular Analysis of Faecal and Duodenal Samples Reveals Significantly Higher Prevalence and Numbers of Pseudomonas Aeruginosa in Irritable Bowel Syndrome. J Med Microbiol. 2011;60(Pt 2):236-45. PubMed PMID: 20947663.
    * Article titles in AMA citation format should be in sentence-case
    TY - JOUR T1 - Molecular analysis of faecal and duodenal samples reveals significantly higher prevalence and numbers of Pseudomonas aeruginosa in irritable bowel syndrome. AU - Kerckhoffs,Angèle P M, AU - Ben-Amor,Kaouther, AU - Samsom,Melvin, AU - van der Rest,Michel E, AU - de Vogel,Joris, AU - Knol,Jan, AU - Akkermans,Louis M A, Y1 - 2010/10/14/ PY - 2010/10/16/entrez PY - 2010/10/16/pubmed PY - 2011/3/2/medline SP - 236 EP - 45 JF - Journal of medical microbiology JO - J. Med. Microbiol. VL - 60 IS - Pt 2 N2 - Intestinal microbiota may play a role in the pathophysiology of irritable bowel syndrome (IBS). In this case-control study, mucosa-associated small intestinal and faecal microbiota of IBS patients and healthy subjects were analysed using molecular-based methods. Duodenal mucosal brush and faecal samples were collected from 37 IBS patients and 20 healthy subjects. The bacterial 16S rRNA gene was amplified and analysed using PCR denaturing gradient gel electrophoresis (DGGE). Pooled average DGGE profiles of all IBS patients and all healthy subjects from both sampling sites were generated and fingerprints of both groups were compared. The DGGE band fragments which were confined to one group were further characterized by sequence analysis. Quantitative real-time PCR (q-PCR) was used to quantify the disease-associated microbiota. Averaged DGGE profiles of both groups were identical for 78.2 % in the small intestinal samples and for 86.25 % in the faecal samples. Cloning and sequencing of the specific bands isolated from small intestinal and faecal DGGE patterns of IBS patients showed that 45.8 % of the clones belonged to the genus Pseudomonas, of which Pseudomonas aeruginosa was the predominant species. q-PCR analysis revealed higher levels (P<0.001) of P. aeruginosa in the small intestine of IBS patients (8.3 %±0.950) than in the small intestine of healthy subjects (0.1 %±0.069). P. aeruginosa was also significantly (P<0.001) more abundant (2.34 %±0.31) in faeces of IBS patients than in faeces of healthy subjects (0.003 %±0.0027). This study shows that P. aeruginosa is detected more frequently and at higher levels in IBS patients than in healthy subjects, suggesting its potential role in the pathophysiology of IBS. SN - 1473-5644 UR - https://www.unboundmedicine.com/medline/citation/20947663/Molecular_analysis_of_faecal_and_duodenal_samples_reveals_significantly_higher_prevalence_and_numbers_of_Pseudomonas_aeruginosa_in_irritable_bowel_syndrome_ L2 - http://jmm.microbiologyresearch.org/pubmed/content/journal/jmm/10.1099/jmm.0.022848-0 DB - PRIME DP - Unbound Medicine ER -