H. pylori infection of the stomach is widespread among human populations including Iranians and is considered to play a major role in the pathogenesis of gastric diseases such as peptic ulcer, adenocarcinoma, and MALT lymphoma. The association between H. pylori virulence markers and disease has been studied in other populations around the world. The aim of this study was to investigate the distribution of H. pylori vacA and cagA genotypes and their association with clinical outcomes in Iran. H. pylori was cultured from gastric biopsy specimens obtained from 137 Iranian patients (58 with duodenal ulcer, 61 with nonulcer dyspeptic [NUD], and 18 with gastric adenocarcinoma). The vacA alleles and cagA genotypes were determined by PCR. The vacA sl allele was present in 107 of the 137 subjects (78%). Multiple strains (m1 and m2) were detected in H. pylori isolates from the patients. VacA s1 genotypes were more frequent in patients with peptic ulcer (46/58; 79%) than in NUD patients (47/61; 77%). CagA was present in 44%of the patients. NUD patients had a frequency of cagA positivity similar to that of the overall population (46%). CagA was present more frequently more than cagA-negative (20%vs. 8%, respectively) in patients with gastric carcinoma (20%) than cagA-negative in patients with gastric carcinoma (8%). This is the first comprehensive study to demonstrate the frequency of colonization with mixed strain, vacA s1, m1 and m2 as the dominant genotype in these Iranian patients, where a high rate of H. pylori infection exists and is similar to the region with a low rate of H. pylori infection. Therefore, host genetics, environmental factors, and the substantial genetic heterogeneity among different H. pylori strains may contribute to the different clinical outcomes. H. pylori infection of the stomach is widespread among human populations including Iranians and is considered to play a major role in the pathogenesis of gastric diseases such as peptic ulcer, adenocarcinoma, and MALT lymphoma. The association between H. pylori virulence markers and disease has been studied in other populations around the world. The aim of this study was to investigate the distribution of H. pylori vacA and cagA genotypes and their association with clinical outcomes in Iran. H. pylori was cultured from gastric biopsy specimens obtained from 137 Iranian patients (58 with duodenal ulcer, 61 with nonulcer dyspeptic [NUD], and 18 with gastric adenocarcinoma). The vacA alleles and cagA genotypes were determined by PCR. The vacA sl allele was present in 107 of the 137 subjects (78%). Multiple (m1 and m2) were detected in H. pylori isolates from the patients. VacA s1 genotypes were more frequent in patients with peptic ulcer (46/58; 79%) than in NUD patients (47/61; 7 7%). CagA was present in 44% of the patients. NUD patients had a frequency of cagA positivity similar to that of the overall population (46%). CagA was present more frequently more than cagA-negative (20% vs. 8 %, respectively) in patients with gastric carcinoma (20%) than cagA-negative in patients with gastric carcinoma (8%). This is the first comprehensive study to demonstrate the frequency of colonization with mixed strain, vacA s1, m1 and m2 as the dominant genotype in these Iranian patients, where a high rate of H. pylori infection exists and is similar to the region with a low rate of H. pylori infection. Thus, host genetics, environmental factors, and the substantial genetic heterogeneity among different H pylori may may contribute to the different clinical outcomes.