Progress in elucidating the pathogenesis of Helicobacter pylori gastric infection and in developing an H. pylori vaccine will be aided by an animal model in which H. pylori can be reliably detected. To validate the use of the mouse model of H. pylori infection, we determined the susceptibility of three inbred strains of mice (C57BL/6J, C57BL/10J and BALB/c) to two VacA+/CagA+ isolates of H. pylori (SPM326 and M1.16) and determined the effectiveness of microbiological, histological and molecular assays for H. pylori detection. For the detection of H. pylori in inoculated mice, reverse transcriptase-polymerase chain reaction was the most sensitive assay (82%), histological evaluation the next most sensitive (66%) and microbiological evaluation the least sensitive (38%); the assays were equally specific (100%). Of the two H. pylori isolates, M1.16 showed the highest rate of colonization, but SPM326 displayed the highest rate of persistent infection. Among the three mouse strains, C57BL/6J mice showed the highest level of both susceptibility to colonization and persistent infection. Anti-H. pylori antibody responses were induced in all inoculated mice and persisted for up to 8 weeks after H. pylori clearance. These results indicate that inbred mice experimentally infected with H. pylori is a reliable model for human infection, but host susceptibility to colonization and persistence of infection are dependent on the H. pylori isolate and the mouse strain.