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Detection of Helicobacter species in liver and stomach tissues of patients with chronic liver diseases using polymerase chain reaction-denaturing gradient gel electrophoresis and immunohistochemistry

Stalke, P; Abu Al-Soud, Waleed LU ; Bielawski, K P; Bakowska, A; Trocha, H; Stepinski, J and Wadström, Torkel LU (2005) In Scandinavian Journal of Gastroenterology 40(9). p.1032-1041
Abstract
Objective. Helicobacter DNA has been detected in the hepatobiliary tree of patients with chronic liver diseases (CLD). The presence of H. pylori in the stomach compared with in the liver of the same patients with CLD has not been studied, therefore to the aim of this study was to investigate the presence of Helicobacter DNA and antigens in the liver and stomach of Polish patients with chronic liver diseases using molecular and immunological methods. Material and methods. Gastric mucosa and liver tissue samples and sera were collected from 97 Polish patients with CLD. Anti-H. pylori antibodies were detected by enzyme immunoassay (EIA), and H. pylori-like antigens detected by immunohistochemistry. Helicobacter DNA was detected in stomach and... (More)
Objective. Helicobacter DNA has been detected in the hepatobiliary tree of patients with chronic liver diseases (CLD). The presence of H. pylori in the stomach compared with in the liver of the same patients with CLD has not been studied, therefore to the aim of this study was to investigate the presence of Helicobacter DNA and antigens in the liver and stomach of Polish patients with chronic liver diseases using molecular and immunological methods. Material and methods. Gastric mucosa and liver tissue samples and sera were collected from 97 Polish patients with CLD. Anti-H. pylori antibodies were detected by enzyme immunoassay (EIA), and H. pylori-like antigens detected by immunohistochemistry. Helicobacter DNA was detected in stomach and liver samples using a semi-nested Helicobacter genus-specific polymerase chain reaction (PCR) assay, and Helicobacter species identified by denaturing gradient gel electrophoresis (DGGE) and sequencing analysis of amplified PCR products. Results. H. pylori was identified by DGGE and sequence analysis in 60/62 (97%) and 25/25 (100%) of the gastric and liver Helicobacter genus-positive samples, respectively, whereas DNA of H. heilmannii was detected in 2/62 (3%) of the Helicobacter genus-positive gastric samples. H. pylori cagA gene was detected in 23/62 (36%) and 3/25 (12%) gastric and liver tissue samples, respectively. H. pylori-like antigens were detected in 61/97 (63%) gastric mucosa and in 40/97 (41%) liver tissue samples. Conclusions. H. pylori - like organisms appeared to dominate the gastric mucosa and liver tissue of Polish patients with CLD. The prevalence of the cagA gene was higher in stomach compared with liver samples, which suggests a possible role of cagA negative H. pylori - like organisms in CLD. On the other hand, no significant correlation was found between the presence of H. pylori - like DNA and antigens in the liver and liver function tests. (Less)
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author
organization
publishing date
type
Contribution to journal
publication status
published
subject
keywords
PCR-DGGE, Helicobacter pylori, DNA sequencing, CagA, chronic liver diseases, 16S rDNA
in
Scandinavian Journal of Gastroenterology
volume
40
issue
9
pages
1032 - 1041
publisher
Taylor & Francis
external identifiers
  • pmid:16165716
  • wos:000231975900006
  • scopus:24744444487
ISSN
1502-7708
DOI
10.1080/00365520510023251
language
English
LU publication?
yes
id
478c7154-12a5-4ac7-bad3-580409042214 (old id 223860)
date added to LUP
2007-08-10 09:30:21
date last changed
2017-10-22 04:50:28
@article{478c7154-12a5-4ac7-bad3-580409042214,
  abstract     = {Objective. Helicobacter DNA has been detected in the hepatobiliary tree of patients with chronic liver diseases (CLD). The presence of H. pylori in the stomach compared with in the liver of the same patients with CLD has not been studied, therefore to the aim of this study was to investigate the presence of Helicobacter DNA and antigens in the liver and stomach of Polish patients with chronic liver diseases using molecular and immunological methods. Material and methods. Gastric mucosa and liver tissue samples and sera were collected from 97 Polish patients with CLD. Anti-H. pylori antibodies were detected by enzyme immunoassay (EIA), and H. pylori-like antigens detected by immunohistochemistry. Helicobacter DNA was detected in stomach and liver samples using a semi-nested Helicobacter genus-specific polymerase chain reaction (PCR) assay, and Helicobacter species identified by denaturing gradient gel electrophoresis (DGGE) and sequencing analysis of amplified PCR products. Results. H. pylori was identified by DGGE and sequence analysis in 60/62 (97%) and 25/25 (100%) of the gastric and liver Helicobacter genus-positive samples, respectively, whereas DNA of H. heilmannii was detected in 2/62 (3%) of the Helicobacter genus-positive gastric samples. H. pylori cagA gene was detected in 23/62 (36%) and 3/25 (12%) gastric and liver tissue samples, respectively. H. pylori-like antigens were detected in 61/97 (63%) gastric mucosa and in 40/97 (41%) liver tissue samples. Conclusions. H. pylori - like organisms appeared to dominate the gastric mucosa and liver tissue of Polish patients with CLD. The prevalence of the cagA gene was higher in stomach compared with liver samples, which suggests a possible role of cagA negative H. pylori - like organisms in CLD. On the other hand, no significant correlation was found between the presence of H. pylori - like DNA and antigens in the liver and liver function tests.},
  author       = {Stalke, P and Abu Al-Soud, Waleed and Bielawski, K P and Bakowska, A and Trocha, H and Stepinski, J and Wadström, Torkel},
  issn         = {1502-7708},
  keyword      = {PCR-DGGE,Helicobacter pylori,DNA sequencing,CagA,chronic liver diseases,16S rDNA},
  language     = {eng},
  number       = {9},
  pages        = {1032--1041},
  publisher    = {Taylor & Francis},
  series       = {Scandinavian Journal of Gastroenterology},
  title        = {Detection of Helicobacter species in liver and stomach tissues of patients with chronic liver diseases using polymerase chain reaction-denaturing gradient gel electrophoresis and immunohistochemistry},
  url          = {http://dx.doi.org/10.1080/00365520510023251},
  volume       = {40},
  year         = {2005},
}