ObjectivesThe objective of this systematic review and meta‐analysis of observational studies was to assess whether herpes simplex virus type 1 (HSV‐1) can infect endodontic periapical lesions.Materials and MethodsStudies with cross‐sectional design investigating HSV‐1 in periapical tissues of patients with symptomatic and asymptomatic acute and chronic apical periodontitis were searched through MEDLINE, Scopus, Embase, Web of Science, and Google Scholar. Pooled HSV‐1 prevalence proportion with 95% confidence interval (95CI) in periapical lesions was assessed with both fixed‐effect and random‐effects models, with/without adjustment for study quality and publication bias. Result robustness was investigated through sensitivity and subgroup analyses.ResultsLiterature search, performed twice, provided 84 items, and eight remained for the meta‐analysis; globally, there were 194 patients mostly adults. The pooled HSV‐1 prevalence proportions, assessed with various methods, were 6.9% (95CI, 3.8–11.3%, fixed‐effect); 6.8% (95CI, 3.6–11.0%, random‐effects); 8.1% (95CI, 4.4–14.5%, quality‐adjusted); and 4.8% (95CI, 2.0–11.4%; adjusted for small‐study effect).ConclusionsThe results indicated that HSV‐1 can colonize the periapical tissues of 3%–11% patients with periapical diseases. Such data do not imply a causative role of HSV‐1 in disease development and advancement. Well‐designed and large‐sized prospective cohort studies should be added in the literature panorama.