liu.seSearch for publications in DiVA
Change search
CiteExportLink to record
Permanent link

Direct link
Cite
Citation style
  • apa
  • harvard1
  • ieee
  • modern-language-association-8th-edition
  • vancouver
  • oxford
  • Other style
More styles
Language
  • de-DE
  • en-GB
  • en-US
  • fi-FI
  • nn-NO
  • nn-NB
  • sv-SE
  • Other locale
More languages
Output format
  • html
  • text
  • asciidoc
  • rtf
Presence of eubacteria in biopsies from Crohn's disease inflammatory lesions as determined by 16S rRNA gene-based PCR
Linköping University, Department of Molecular and Clinical Medicine, Clinical Microbiology. Linköping University, Faculty of Health Sciences.
Linköping University, Department of Biomedicine and Surgery, Surgery. Linköping University, Faculty of Health Sciences.
Linköping University, Department of Biomedicine and Surgery, Surgery. Linköping University, Faculty of Health Sciences.
Östergötlands Läns Landsting, Centre for Laboratory Medicine, Department of Clinical Microbiology. Linköping University, Faculty of Health Sciences.
Show others and affiliations
1999 (English)In: Journal of Medical Microbiology, ISSN 0022-2615, Vol. 48, no 3, 263-268 p.Article in journal (Refereed) Published
Abstract [en]

The aim of this study was to search for putative microbial agents in Crohn's disease (CD) tissues by bacterial broad-range 16S rDNA PCR combined with genus- and species-specific DNA hybridisation analysis. Biopsies taken both surgically and endoscopically from the terminal ileum of 11 CD patients and 11 control patients were investigated. Significant amounts of eubacteria were demonstrated in biopsies taken endoscopically from both affected and unaffected individuals; the biopsies taken at surgery from control patients were negative. Three of five biopsies taken surgically from CD patients harboured Helicobacter spp.-, Mycobacterium paratuberculosis-, Listeria monocytogenes- and Escherichia coli-like 16S rDNA sequences. These findings show the importance of the sampling method chosen when combined with molecular typing of eubacteria in intestinal tissues. The mixed bacterial flora found in the surgical biopsies from CD patients supports the idea that the enteric microflora enters primary lesions where secondary bacterial colonisers may elicit a chronic inflammatory syndrome.

Place, publisher, year, edition, pages
1999. Vol. 48, no 3, 263-268 p.
National Category
Medical and Health Sciences
Identifiers
URN: urn:nbn:se:liu:diva-25305DOI: 10.1099/00222615-48-3-263PubMedID: 10334593Local ID: 9746OAI: oai:DiVA.org:liu-25305DiVA: diva2:245633
Available from: 2009-10-07 Created: 2009-10-07 Last updated: 2012-08-01
In thesis
1. Identification and characterisation of bacteria based on 16S rDNA techniques with special reference to Helicobacter pylori in the gastro-intestinal tract
Open this publication in new window or tab >>Identification and characterisation of bacteria based on 16S rDNA techniques with special reference to Helicobacter pylori in the gastro-intestinal tract
2000 (English)Doctoral thesis, comprehensive summary (Other academic)
Abstract [en]

The overall aim of this study was to establish molecular techniques for the detection and identification of "difficult to grow" - bacteria in mixed bacterial populations in clinical samples without the need for culturing procedures.

Material and Methods: Thirty-nine strains of Mobiluncus were used as a model system for phylogenetic classification of fastidious bacteria based on 16S ribosomal DNA (rDNA) sequences. To test the application of the 16S rDNA broad range PCR concept for detecting bacteria clinically, urine samples spiked with Chlamydia trachomatis elementary bodies and real urine test samples from 12 C. trachomatis positive and negative male volunteers were tested in a semiblind manner against routine procedures. Furthermore, gastric biopsy samples from 22 individuals (13 defined as having Helicobacter pylori-associated gastritis, and 9 defined as normal controls) were evaluated for the presence of Helicobacter 16S rDNA and the virulence genes cagA, vacA, and ureA. PCR products were also applied to temporal temperature gel electrophoresis (TTGE) gels for profiling the microbial flora. Finally, intestinal biopsies from 22 patients (11 diagnosed as Crohn's disease (CD), and 11 non-CD patients) were investigated using probes targeting potential pathogens that have been suggested to be involved in CD.

Results: We were able to confirm the current species designation of Mobiluncus, although the results did not support the division of M. curtisii into subspecies. For the detection of C. trachomatis in urine, the in-house system was shown to be as sensitive as a commercially available PCR system. The search for Helicobacter pylori in gastric biopsies revealed the presence of Helicobacter DNA in 20 of 22 individuals. The molecular techniques were apparently too sensitive compared with routinely used techniques. TTGE revealed a complex microbial flora both in the normal control group and in the gastritis group, with dominance of Helicobacter in the gastritis group. The present results might lend support to the hypothesis that Helicobacter are indigenous biota of the human stomach. cagA was amplified in all Helicobacter positive specimens. None of the specimens in the control group carried a H. pylori type strain identical vacA genotype. ureA negative mutants were also found in this group. The 16S rDNA sequence data might also indicate phylogenetic heterogeneity. The mixed bacterial flora found in CD inflammatory lesions is consistent with the idea that the enteric microflora enters primary lesions, where secondary invaders may elicit chronic inflannnatory response.

Conclusion: This thesis has demonstrated the usefulness of 16S rDNA based techniques for detection, identification, and characterisation of individual bacterial pathogens as well as for profiling of mixed bacterial flora in clinical specimens.

Place, publisher, year, edition, pages
Linköping: Linköpings universitet, 2000. 112 p.
Series
Linköping University Medical Dissertations, ISSN 0345-0082 ; 622
National Category
Medical and Health Sciences
Identifiers
urn:nbn:se:liu:diva-28646 (URN)13802 (Local ID)91-7219-579-7 (ISBN)13802 (Archive number)13802 (OAI)
Public defence
2000-04-28, Berzeliussalen, Hälsouniversitetet, Linköping, 09:00 (Swedish)
Opponent
Available from: 2009-10-09 Created: 2009-10-09 Last updated: 2012-08-01Bibliographically approved

Open Access in DiVA

No full text

Other links

Publisher's full textPubMed

Authority records BETA

Söderholm, Johan D.Olaison, GunnarJonasson, JonMonstein, Hans-Jürg

Search in DiVA

By author/editor
Söderholm, Johan D.Olaison, GunnarJonasson, JonMonstein, Hans-Jürg
By organisation
Clinical MicrobiologyFaculty of Health SciencesSurgeryDepartment of Clinical Microbiology
In the same journal
Journal of Medical Microbiology
Medical and Health Sciences

Search outside of DiVA

GoogleGoogle Scholar

doi
pubmed
urn-nbn

Altmetric score

doi
pubmed
urn-nbn
Total: 77 hits
CiteExportLink to record
Permanent link

Direct link
Cite
Citation style
  • apa
  • harvard1
  • ieee
  • modern-language-association-8th-edition
  • vancouver
  • oxford
  • Other style
More styles
Language
  • de-DE
  • en-GB
  • en-US
  • fi-FI
  • nn-NO
  • nn-NB
  • sv-SE
  • Other locale
More languages
Output format
  • html
  • text
  • asciidoc
  • rtf