The aims of the study were to evaluate the diagnostic accuracy of a rapid whole blood test (one-step BM-Test Helicobacter pylori) in different blood collections and compare it with a quantitative ELISA test (HEL-p TEST II). One hundred four dyspeptic patients were studied, and 53 (51%) were H. pylori-positive as determined by [13C] urea breath test. The area under the ROC curve was higher for HEL-pTEST II compared to the one-step BM-Test with either fingerstick or venipuncture (0.972 vs 0.721 and 0.714, P = 0.000). The most appropriate cutoff reading time was 5 min for the one-step BM-Test. The sensitivity, specificity, and accuracy were 52.8%, 88.2%, 70.2% for fingerstick; 62.3%, 80.4%, 71.2% for venipuncture; and 90.6%, 92.2%, 91.3% for HEL-pTEST II. We conclude that there is no difference in diagnostic performance between capillary and venous blood for rapid whole blood test, and extending the reading time beyond the manufacturer's suggestion for the one step BM-Test does not improve its accuracy.