The presence of neutralising antibodies (NAbs) against adenovirus in the population is a major hurdle preventing the effective use of replication-defective adenoviruses (Ads) as candidates for gene therapy and vaccine vectors for many diseases. Only a few studies have described the epidemiology of pre-existing immunity to chimpanzee Ads in China. To assess the prevalence of NAbs to chimpanzee adenovirus serotypes 6 and 7 (AdC6 and AdC7), we enrolled 998 healthy participants from five regions in China as well as 196 chronic hepatitis B virus (HBV) patients and 193 primary hepatocellular carcinoma (HCC) patients from Chongqing, China. The total seroprevalence rates of AdC6 and AdC7 NAbs in the healthy participants were 12.22 % (122/998) (95 % confidence interval [CI], 10.34-14.40 %) and 13.13 % (131/998) (95 % CI, 11.17-15.36 %), respectively. The seroprevalence rates of AdC6 and AdC7 NAbs in the HBV patients were 21.43 % (42/196) (95 % CI, 16.26-27.69 %) and 25.51 % (50/196) (95 % CI, 19.92-32.04 %), respectively. The seroprevalence rates of AdC6 and AdC7 NAbs in the HCC patients were 27.46 % (53/193) (95 % CI, 21.65-34.15 %) and 31.09 % (60/193) (95 % CI, 24.98-37.93 %), respectively. The seroprevalence rates of these Ads were not associated with age and gender. The present study may provide useful insights for developing future AdC-based vaccines and gene therapies.