Abstract Background: Helicobacter pylori (H. pylori) is one of the most common chronic bacterial infections in the world, and it is currently estimated that approximately half of the world's population is infected with the bacterium. The correct diagnosis and effective treatment of H. pylori gastric infection are essential in controlling this condition. The available diagnostic methods have advantages and limitations related to factors such as age of patients, technical difficulty level, costs and extensive accessibility in hospitals. The eradication therapy of H. pylori infection is still a challenge for gastroenterologists. One of the main causes of failure in H. pylori eradication is antibiotic resistance. Biopsy cultures are the most widely used methods among the antimicrobial susceptibility tests. Aim of Study: The aim of this work was to determine the proper regiment of H. Pylori treatment according to gastro-duodenal tissue culture and antibiotic sensitivity. Patients and Methods: This study included 50 patients with Helicobacter Pylori Ag positive stools and positive occult blood in stool. Their age ranged from 1.5 to 17 years old, 60% of them were female. 37 patients showed positive gastric and duodenal cultures and received treatment according to results of cultures. Results: Antibiotic sensitivity of gastric culture shows that Levofloxacin had the highest antibiotic sensitivity (93.5%), followed by clarithromycin and amoxicillin in (77%) and (67.7%) respectively. But, 100% of cases were resistant to the Metronidazole. In duodenal culture, Levofloxacin had the highest antibiotic sensitivity (96.4%), followed by, clarithro-mycin and amoxicillin in (71.4%) for each. But, 100% of cases were resistant to the Metronidazole. All patientsreceived PPI and Levofloxacin, while about 59% and 30% of patients received Amoxacillin and Clarithromycin respectively. Conclusion: Treatment of Helicobacter Pylori should be tailored according to tissue culture and sensitivity.