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Helicobacter pylori (HP) infection is associated with significant gastric mucosal inflammation. We aimed to determine the clinicopathologic features associated with HP in gastric biopsy.Three hundred ninety-six gastric biopsies were evaluated including 165 HP-positive cases and 231 randomly selected HP-negative controls. HP was detected using hematoxylin and eosin (H&E), Giemsa, and immunohistochemistry staining. The univariate and multivariate analyses were conducted to study the relationship of clinicopathologic characteristics and HP infection. Among the HP-positive cases, 131 cases were confirmed by H&E staining and 34 cases were confirmed by Giemsa or immunohistochemistry staining. Compared with chronic inactive gastritis, chronic active gastritis was more likely associated with having HP detected by H&E. Males were more likely to have HP gastritis than females (odds ratio: 1.72, P=0.01). The patients who had chronic active gastritis or chronic gastritis (moderate or severe) were more likely to have HP infection than patients with mild chronic gastritis (P<0.001). Conversely, patients who had intestinal metaplasia were less likely to have HP infection than patients without intestinal metaplasia (odds ratio: 0.22, P<0.001). However, concurrent atrophic gastritis was not related to HP infection (P=0.37). HP infection history was not associated with HP infection (P=0.74).HP detection in gastric biopsies is associated with active inflammation, male sex, and the lack of intestinal metaplasia, but not atrophic gastritis or HP infection history. Routine ancillary staining may not be required for HP detection in all biopsy specimens. We do not recommend ancillary staining for mild chronic inactive gastritis.