The relief from clinical symptoms of Helicobacter pylori infection caused by Lactobacillus acidophilus treatment was analyzed by a 13C-urea breath test. To this end, 200 patients diagnosed with Helicobacter pylori infection were divided into two equal groups of 100 patients, designated as the control and study groups, respectively. The control group received conventional quadruple therapy that included enteric-coated esomeprazole magnesium tablets, amoxicillin, clarithromycin, and colloidal pectin bismuth for 2 weeks. The study group, in addition to the conventional quadruple therapy, received Lactobacillus acidophilus tablets orally for 2 weeks. The Helicobacter pylori eradication rate (P < 0.05) and total effective rate (P < 0.05) of the study group were higher than the control group. The incidence of adverse reactions including acid reflux, upper abdom¬inal pain, abdominal fullness, and vomiting in the study group was lower (P < 0.05) than the control group. There was no statistically significant difference (P > 0.05) in the levels of tumor nuclear factor-α, high sensitivity C-reaction protein, and interlukin-6 between two groups before treatment. However, the level of these indicators in the study group after treatment was lower than that in the control group. Similarly, there was no statistically significant difference in CD3+, CD4+, CD8+, or CD4+/CD8+ between the two groups before treatment. However, the level of CD3+, CD4+, and CD4+/CD8+ after treatment was significantly higher, while CD8+ was significantly lower in the study group. In conclusion, Lactobacillus acidophilus treat¬ment exhibits a high eradication rate of Helicobacter pylori with good safety and relief from Helicobacter pylori-associated clinical symptoms.