Objective To investigate the prevalence of hepatitis B virus (HBV) and hepatitis C virus (HCV) in human immunodeficiency virus (HIV)-positive patients infected by blood (with exception of intravenous drug users) and sexual transmission routes in China. Methods Three hundred and sixty-two cases of HIV positive patients from 13 acquired immune deficiency syndrome (AIDS) centers of China (those intravenous drug users were excluded) were analyzed retrospectively. The serological markers of HBV infection including HBsAg, anti-HBs, HBeAg, anti-Hbe, anti-HBc and anti-HCV antibodies were detected by the chemiluminescence immunoassay(CMIA) or enzyme-linked immunosorbent assay (ELISA). Numeration and measurement data were hnalyzed using chi square test and t test, respectively. Results Both HBV markers and HCV antibody were detected in 315 HIV postitive patients. Among them, fourteen cases were HBsAg positive, with the percentage of 4.4%. One hundred and fifty-eight cases (50. 2%) were anti-HCV antibody postive and 157 cases (49.8%) were negative. Only 2 cases (0.6%) were H1V, HBV, HCV co-infection. In the anti-HCV positive group, the percentage of blood and sexual transmission were 92 % and 4 %, respectively, with a predominant blood transmission; while in the anti-HCV negative group, the percentage of blood and sexual transmission were 11% and 66% respectively, dominating with sexual transmission. The duration of HIV infection, CD4+ T cell absolute numbers, alanine aminotransferase (ALT) and aspartate aminotransferase (AST) levels were all higher in the anti-HCV positive group than those in the anti-HCV negative group. Meanwhile, there were 2 cases (1.3%) of HBsAg positive in the anti-HCV positive group while 12 cases (7.6%) in the anti-HCV negative group, with a statistically significant difference (X2= 7.542, P<0.01). HBV DNA detection was performed in 10 HBsAg positive patients, four of them were HBV DNA positive but all were anti-HCV negative. Fifty-seven patients with anti-HCV positive received HCV RNA detection and 63.2% of them were positive. Conclusions HBV and HCV co-infection exists in HIV-infected Chinese patients, predominantly with HCV co-infection and by blood transmission. HCV infection may aggravate the liver injury of HIV infection and, at the same time, may have some mechanisms to interfere HBV replication.