Background: Human cytomegalovirus (HCMV) appears to be more frequent in periodontally affected patients than in healthy control groups. Based on this assumption, it has been suggested that HCMV may play a role in the pathogenesis of periodontal disease. Objective: The objective of this uncontrolled study was to assess the occurrence of HCMV in a large unselected population of periodontally healthy subjects. Study Design: Fifty consecutive periodontally healthy patients satisfied the inclusion criteria. Two samples of gingival crevicular fluids were taken from two non-bleeding on probing sites for each patient. Samples were collected from the anterior and the posterior area. Polymerase chain reaction (PCR) was used to identify the presence of HCMV. Results: HCMV was detected in 17 (33%) out of 50 participants. Ten subjects showed presence of HCMV on both anterior and posterior sites, whereas the remaining 7 only had HCMV present in the anterior sites. No differences were noticed between HCMV positive and HCMV negative in terms of smoking (p = 0.33), drinking habits (p=0,94) or the presence of prosthodontic restorations (p= 0,89). Conclusions: HCMV was detected in a high proportion of periodontally healthy subjects. Its presence was not found to be influenced by smoking or drinking habits.