Introduction: Anti-hepatitis B core antibody (Anti-HBc) alone is defined as the presence of anti-HBc in the absence of HBsAg and anti-HBs. The significance of this serological pattern as a predicting factor for occult hepatitis B virus (HBV) infection remains largely unknown. This study aimed to assess the significance of anti-HBc alone in predicting occult HBV infection in high-risk and low-risk individuals. Methodology: A total of 926 individuals were enrolled in this study, including 289 hemodialysis (HD) and 106 HIV-infected patients who were considered as a high-risk group and 531 blood donors who were considered as low-risk. HBsAg, anti-HBs, anti-HBc were tested in all subjects. The presence of HBV-DNA was determined quantitatively in patients with anti-HBc alone by real-time PCR. Results: Of the 395 high-risk patients, 40 cases (10.13%) had anti-HBc alone, while 11 subjects (2.07%) out of 531 blood donors had anti-HBc alone. HBV-DNA was detected in 12 out of 40 (30%) high-risk patients and none of the blood donors with anti-HBc alone. Conclusion: Our study showed that the serological pattern of anti-HBc alone could reflect occult HBV infection in high risk cases but did not presume occult HBV infection in low-risk individuals.