Introduction: Helicobacter pylori infection is associated with gastritis, peptic ulcer, and gastric cancer. We conducted a cross-sectional study to compare five diagnostic tests for H. pylori infection and studied the epidemiology of the infection in Bangladesh. Methodology: Bangladeshi patients with dyspeptic symptoms referred for endoscopic examination were enrolled in this study. Each patient underwent upper endoscopic examination and four gastric biopsy specimens were taken. We used 5 tests for the diagnosis of H. pylori; culture, histology confirmed by immunohistochemistry, rapid urease test (RUT), urinary and serological test. Demographic and environmental variables were collected. Results: A total of 133 patients participated in the study, 61 males and mean age 37.3 ± 12.3 years. We used the culture and/or histology results as the gold standard to estimate the sensitivity, specificity, positive and negative predictive values for the studied diagnostic tests. RUT, culture and histology had high sensitivity and specificity with moderate positive and negative likelihood ratio, whereas urine test and serology showed a good sensitivity and specificity but poor likelihood ratio. The overall prevalence of H. pylori among study subjects was 47% with no difference between gender and age groups. Conclusions: The invasive tests showed better performance than noninvasive tests among Bangladeshi population. The overall prevalence of H. pylori was less than the previously reported in the region with no difference among all age groups.