OBJECTIVE To assess the seroprevalence of Helicobacter pylori infection in a large series of patients with upper gastrointestinal symptoms referred for their first upper gastrointestinal endoscopy, and to evaluate any association with disease. METHODS A large survey of 3281 dyspeptic patients undergoing their first endoscopic examination was undertaken, involving 93 centres in Italy. A blood sample was taken from each participant, for measuring IgG antibodies against H. pylori using a commercially available kit, and two biopsies of the antral gastric mucosa were obtained for evaluating a Giemsa-stained specimen. RESULTS Endoscopic macroscopic diagnoses included normal mucosa (25.3%), gustroduodenitis (51.6%), gastric and duodenal ulcers (3.7 and 14.9%, respectively) and other conditions, including gastric cancer (0.8%). Overall, the seroprevalence result was 71.3% with a strong positive association with increasing age and male sex and a negative one with educational level. According to endoscopic diagnoses, the association with H. pylori seropositivity was highest for duodenal and gastric ulcer (multivariate odds ratio: 6.1 and 2.2) and lowest for carcinoma. The comparison between the results of serology and the single Giemsa-stained specimen showed good reliability of H. pylori IgG, particularly in a subgroup (n = 2056) for which the interpretation was performed by a single dedicated pathologist: sensitivity and specificity were 92 and 78%, respectively. CONCLUSIONS Commercial IgG serology is a reliable tool for the assessment of H. pylori infection in large-scale multicentre surveys. A very high seroprevalence among dyspeptic patients was confirmed, particularly in the presence of peptic disease. Factors associated with the infection were very similar to those usually reported in the general population, but male patients showed a significantly higher prevalence.