Cytology and the rapid urease test on gastric biopsies may diagnose Helicobacter pylori (H. pylori) infection within an hour. We evaluated the sensitivity and reproducibility of touch cytology (imprints from biopsies). In 19 patients with duodenal ulcer, biopsies were obtained from the antrum, fundus, and bulb. H. pylori was diagnosed in 42 sites on smears: 28 by culture and 23 by histology. H. pylori was present in the antrum and fundus in 16 and in the bulb in 11. Assessment of paired antral biopsies in 29 additional patients with or without touch cytology (imprints) before specimens were sent for histology or culture revealed no difference for the presence of H. pylori. A second reading of the 58 smears by a second observer revealed agreement on the presence or absence of H. pylori in 53 (91%). In conclusion, touch cytology is a simple rapid, sensitive, and reproducible diagnostic method for H. pylori that does not alter the quality of biopsies for subsequent culture or histologic examination. For the first time, diagnostic methods have been compared on the same biopsies, eliminating sampling variation.