The hygienic quality of milk has an important public health importance. Helicobacter pylori has a foodborne route and especially food with animal origin. This present study was carried out in order to isolation the H. pylori from cow milk samples using culture and PCR method. A total of 120 bovine milk and 120 dairy product samples were collected from the supermarkets of various parts of Iran. The animals whose milk samples collected for this study were clinically healthy.All samples were cultured and those that were H. pylori-positive were subjected for PCR method fordetection of the H. pyloriure C gene.Of 240 samples studied, 33 (13.75%) were positive for the H. pylori using the culture method. Raw bovine milk were the most contaminated (16.66%) but traditional cream were the less contaminated (7.5%) samples. Significant differences was seen for the prevalence of H. pylori between raw bovine milk and traditional cream samples (P =0.027). All of the positive colonies of H. pylori were confirmed using the ureC gene based-PCR method. This study showed that cow milk and traditional dairy product samples are the sources of H. pylori infection for humans.