Tenofovir disoproxil fumarate (Tenofovir DF) is a nucleotide analogue. This multicentre study reports retrospectively the long-term efficacy and safety data with tenofovir DF treatment in nucleosid(t)e-naive, hepatitis B e antigen (HBeAg)-positive chronic hepatitis B patients. Thirty-one patients (11 females, 20 males) received 245 mg tenofovir DF per diem. All patients' initial serum hepatitis B virus (HBV) DNA levels were over 2,000 IU/ml. Serum alanine aminotransferase (ALT) levels, HBeAg, hepatitis B e antibodies (anti-HBe), hepatitis B surface antigen (HBsAg), hepatitis B surface antibodies (Anti-HBs), HBV DNA, creatinine and urea levels were evaluated at baseline, and at weeks 12, 24, 48 and 96 during therapy. Thirty-one patients completed 96 weeks of treatment. Mean age was 37.6 ± 9.4 years. The initial mean value of ALT was 79 ± 39.9 IU/L. At baseline, mean of fibrosis (Ishak) of liver biopsies was 2.3 ± 0.7. Two of the patients (5.9%) achieved HBV DNA less than 300 copies at week 12 of treatment and 97.1 % at week 96. HBeAg loss was observed in 6.7% of patients. At week 96, HBsAg loss was not observed in any of the patients. Mean ALT at week 48 was 32.7 U/L, at week 96 32.6 U/L. Renal safety was good. Creatinine remained stable. Tenofovir DF was well tolerated and produced potent, continuous viral suppression with increasing HBeAg loss.