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Gene therapy has been considered to be the promising and accomplishing approach to counter persistent HBV infection and primary hepatocellular carcinoma(HCC).Replication-defective adenoviruses were one of the most potent vectors for targeted gene delivery.However,clinical application to the best-studied human adenovirus type-5(AdHu5)is limited by the high prevalence of pre-existing neutralizing antibodies(NAb)due to natural infection,while rare Ad serotypes such as chimpanzee adenovirus type-68(AdC68)and human adenovirus type-26(AdHu26)could circumvent pre-existing immunity and serve as substitutes for AdHu5.We collected blood samples from 196 patients with chronic HBV infection and 193 patients with HCC in China,the highly epidemiological area with HBV infection,to assess NAb levels for AdC68,AdHu26 and AdHu5.The seroprevalence rate ofAdHu5,AdHu26 and AdC68 neutralizing antibodies for patients with chronic hepatitis B was 84.96%(166/196),58.16%(114/196)and 39.80%(78/196),respectively.And among patients with HCC,the incidence was 66.30%,67.4% and 32.1% for AdHu5,AdHu26 and AdC68,respectively.More than 90% of the AdC68 Nab titers was less than 40 in both groups.Our investigation suggests that chimpanzee-derived vector AdC68 may have the advantage for therapeutic gene delivery in Chinese patients with chronic hepatitis B or HCC.