DNA Variation of Helicobacter Pylori in the Gastroduodenal Disease

Roe, Im Hwan; Kim, Chang In; Ha, Dong Ryul; Jin, Young Joo; Song, Il Han; Lim, Chang Young; Kim, Jung Won; Kim, Jung Taik; Kim, Jong Hwa; Yeom, Jung Sun

Abstract

BACKGROUND: The evidence for H. pylori as a gastrointestnal pathogen is now very strong, if not overwhelming. Among the pathogenic factors of H. pylori, flagella and urease are considered to be major factors causing the gastrododenal disease. We observed the gene diversity of H. pylori using the PCR-amplified 1.4Kb fla A gene and 0.9Kb ure B gene and examined the relationship between the gene pattern and the gastroduodenal disease. METHOD: Fifty-one cases of isolated strains were cultured at the Helicobacter-selective blood agar plates. To compare the gene diversity among the isolates of gastroduodenal disease genotypes was analyzed by PCR-based RFLP. 1.4Kb fla A gene and 0.9Kb ure B genes from isolates were amplified by PCR and digested with Hae 3 restriction enzymes to observe the restriction fragment length polymophysm. Protein patterns were also compared to examine the antigenic variations. Total cell proteins, and octyl-glucose extracts from isolates were analyzed by SDS-PAGE gel electrophoresis.
RESULTS
41 cases (80.4%) of H. pylori were isolated in the 51 cases of gastroduodenal diseases. We could classify theses isolates 3 types of PCR-RFLP in the fla A gene, 900+500bp, 500+500+400bp, 600+800bp, and 9 types in the ure B gene. PCR-RFLP in the fla A gene and ure B gene of the isolates was different from the standard strain of Australia and the genetic diversity was not related to the types of the gastroduodenal disease. We demonstrated variations in the protein pattern and antigenic profiles among the isolates by SDS-PAGE analysis. These data also did not show any relationship between protein pattern and types of gastroduodenal diseases. CONCLUSION: Tese studies showed many different gene diversity in the flagella and urease gene without any relationship with the types of gastoduodenal disease. And variable protein pattern were noted among the strains of H. pylori. Further studies to demonstrate the pathgenecity of H. pylori should be continued even if there was no relationship between the genomic diversity of the flagella or urease and the types of gastroduodenal disease.