In general, cytokines encoded by different genes of human genome might strongly influence host cell-mediated immune responses, which play an important role in the clearance of virus by the infected host. Interferon gamma (IFN-gamma) produced by T lymphocytes and natural killer cells plays an essential role in affecting cellular immune responses. A functional study demonstrated that two single nucleotide polymorphisms located in the IFN-gamma gene intron (at positions +874 and +2109) were involved in its transcriptional regulation. The aim of this study was to evaluate whether IFN-gamma gene polymorphisms or its haplotypes might be associated with predisposition to hepatitis B virus (HBV) infection in the Chinese population. The study included 181 cases with HBV infection and 272 gender, age-matched healthy controls. All genotyping were identified by polymerase chain reaction in association with the measurement of amplification refractory mutation system. A significant difference was observed between case and control groups. The frequency of +874A allele was significantly higher in patients than in controls (OR = 2.25, 95%CI = 1.69-2.99, P < 0.0001). However, no significant difference was found in the allelic frequencies of IFN-gamma +2109A/G between cases and controls (P > 0.05). By haplotype analysis, the frequency of haplotype AG (+874A and +2109G) revealed a significant difference in the cases in comparison to controls (P < 0.0001). Multiple logistic regression analysis showed that individuals possessing haplotype AG had an increased likelihood of HBV infection (OR = 8.14, 95%CI = 4.98-13.30). Our results suggest that haplotype AG containing +874A and +2109G may be a crucial risk factor of genetic susceptibility to HBV infection in the Chinese population.