Abstract
BACKGROUND: The unique survival environment of Helicobacter pylori (H. pylori) presents challenges in establishing appropriate in vitro culture conditions. The fastidious nature of H. pylori and its relatively low growth rate emphasize the need for customized culture conditions, which are essential for both research and clinical applications. AIM: To identify the optimal liquid culture medium for H. pylori using both reference and clinical strains. METHODS: Nine H. pylori strains were incubated in 10 different broth media commonly used for bacterial cultures under microaerophilic conditions. Bacterial concentrations were estimated using serial dilutions and the pour plate method. A range of H. pylori initial inoculum concentrations was tested for each strain. For growth evaluation, optical density at 600 nm was measured at 24, 48, and 72 hours. The growth trends of H. pylori were compared among strains and media. RESULTS: All H. pylori strains grew successfully over time, regardless of the initial inoculum concentration. The specific growth trends and the broth that yielded the highest optical density value at each time point varied with strain. The reference strains (H. pylori 26695, ATCC 43504, and J99) consistently exhibited robust growth, whereas the clinical strains displayed distinct media preferences. Overall, bacterial growth was higher in chopped meat carbohydrate broth, Columbia broth, and fastidious anaerobe broth than in Brucella broth after 72 hours. CONCLUSION: Chopped meat carbohydrate broth was the most effective for H. pylori liquid culture, and Columbia broth and fastidious anaerobe broth also supported greater growth than Brucella broth.