Helicobacter pylori, a gastric pathogen that colonizes approximately 50% of the world’s population. Infection with H.pylori is the strongest known risk factor for some GI disorders including gastric cancer. Probiotics have considerable potential for preventive or therapeutic applications in various gastrointestinal disorders. So, the aim of this study was in vitro study of the potential inhibitory effect of Lactobacillus strains on H.pylori Growth. The aim of the potential inhibitory effect of Lactobacillus strains on H.pylori by using a standard antimicrobial plate well diffusion assay and liquid media assay. Among the two, L. acidophilus showed maximum zone of inhibition of a diameter of 22 18 mm against H.pylori. When the supernatant of the culture media of L. acidophilus was diluted with PBS, the zone of inhibition was decreased. On the other hand, L. rhamnosus showed maximum zone of inhibition of 23 mm in diameter against H.pylori. Unlike L. acidophilus, supernatant of the culture media of L. rhamnosus did not show any zone of inhibition upon dilution with PBS. Bacteriocins from Lactobacillus strains were found to be heat-stable (1210C for 15 min) and active over a wide pH range of 5.0-9.0. Addition of surfactants (EDTA, SDS, CTAB) up to 1% to crude supernatant containing Bacteriocins showed increase in antibacterial activity; whereas NaCl concentration of 2% increased the antibacterial activity. Study concluded that the growth of the cancer bacteria H.pylori can be controlled by Lactobacillus strain but still requires optimization.