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Globally, the upscaling prevalence of Helicobacter pylori infection has become a huge concern. This organism is a microaerophilic gram-negative bacterium that infects the stomach and duodenum of humans and has been rewarded as a class 1 carcinogen. Approximately 50% of the world populars are known to be infected with Helicobacter pylori. This study evaluated the knowledge as well as the prevalence of Helicobacter pylori among the students of colleges of Medicine and faculty of Pharmacy, University of Benin. A total of 107 who fulfilled the inclusion criteria were recruited for the study. Ethical clearance and informed consent were retrieved and a well-structured questionnaire was administered to each participant. Whole blood samples were collected from the participants and screened for Helicobacter pylori antibody using rapid test kits (Rostec). The information obtained was analyzed using Stata version 16. P-values <0.05 were considered as statistically significant. Out of the 107 students tested, 31 (29.0%) were positive for H. pylori. The prevalence was higher in females 24 (77.4%) as against 7 (22.6%) in males. The students within the age group of 21-25 years reported the highest prevalence. There was no statistical significance in the prevalence of Helicobacter pylori between the age groups (P=0.316) and gender groups (P=0.981). There was a statistical significance in the prevalence of H. pylori in accomodation type (P=0.014). The purpose of this work was to generate information focusing on the prevalence of Helicobacter pylori as well as their knowledge. Information generated will help formulate health interventions needed to check these problems and offer preventive approach to the students of the College of Medicine and Faculty of Pharmacy as well as the general population of University of Benin.