Objectives: Health care workers, including dentists, are at the front line for acquiring blood-borne virus infections. This study aimed to investigate the prevalence of hepatitis B, hepatitis C and human immunodeficiency viruses among Iranian dentists. Material and Methods: The survey included 1628 dental health care workers who attended the 51st annual Congress of the Iranian Dental Association. Data on the risk of blood-borne virus transmission during health care and infection control practices were gathered from self-administered questionnaires. Sera were screened serologically by enzyme-linked immunosorbent assay (ELISA). The positive samples were examined by polymerase chain reaction (PRC) followed by direct sequencing. Results: Six (0.36%) and 81 (5.0%) were positive for hepatitis B surface antigen (HBsAg) and (anti-hepatitis B virus core antigen (anti-HBc), respectively. Only 1 (0.061%) was positive for anti-hepatitis C virus (anti-HCV). No case was positive for anti-human immunodeficiency virus (anti-HIV). One case was diagnosed as being occult hepatitis B virus (HBV) infection. One thousand five hundred thirty-five (94.3%) of participants had received at least 1 dose of HBV vaccine. One thousand three hundred fifty-nine (88.5%) contained hepatitis B surface antibody (anti-HBs) > 10 IU/ml, of whom 55 (4.0%) were anti-HBc positive, suggesting that they had been infected with HBV in the past. Anti-HBc positive cases had past histories of hepatitis, either their own or their spouses’. Individuals with inadequate anti-HBs levels (< 10 IU/ml) were significantly more prevalent among anti-HBc positive cases (p < 0.001). Conclusions: The prevalence of blood-borne viruses among dental HCWs found in this study was lower than past reports from Iranian dentists and general population. The implementation of HBV vaccination together with improvement in infection control procedures has reduced the potential for risk infection among Iranian dentists.