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.
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.


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.
Stalke
Piotr
P
Department of Infectious Diseases, Medical University of Gdansk, Gdansk, Poland. pstalke@amg.gda.pl
Al-Soud
Waleed Abu
WA
Bielawski
Krzysztof P
KP
Bakowska
Alicja
A
Trocha
Hanna
H
Stepinski
Jan
J
Wadström
Torkel
T
D000293:Adolescent; D000328:Adult; D000368:Aged; D000907:Antibodies, Bacterial / Q000032:analysis; D000942:Antigens, Bacterial / Q000032:analysis; D001707:Biopsy, Needle; D004269:DNA, Bacterial / Q000032:analysis*; D004591:Electrophoresis, Polyacrylamide Gel; D005260:Female; D005753:Gastric Mucosa / Q000382:microbiology* / Q000473:pathology; D005773:Gastroscopy; D016480:Helicobacter pylori / Q000235:genetics* / Q000276:immunology / Q000302:isolation & purification; D006801:Humans; D007124:Immunoenzyme Techniques; D007150:Immunohistochemistry / Q000379:methods*; D008099:Liver / Q000382:microbiology* / Q000473:pathology; D008107:Liver Diseases / Q000382:microbiology* / Q000473:pathology; D008297:Male; D008875:Middle Aged; D016133:Polymerase Chain Reaction; D012189:Retrospective Studies
D000907:Antibodies, Bacterial; D000942:Antigens, Bacterial; D004269:DNA, Bacterial