Abstract: Objective To study the influence of hemolysis and lipoidemia on real-time fluorescence quantitative polymerase chain reaction (RT-PCR) of HBV DNA. Methods (1)Clinical HBV DNA specimens were collected, and were divided into level I and level II according to HBV DNA concentrations. HBV DNA negative but severe blood lipid samples and normal blood samples were collected. These specimens were mixed in different proportions and divided into group A, B, C by different TG concentrations. (2)Clinical HBV DNA specimens were collected, three tubes of blood were obtained from each person, and two of them were frozen in a-20 ℃ refrigerator for different lengths of time to make hemolytic samples of different degrees. Then, they were divided into group D, E, F according to the degree of hemolysis. Results There is no statistically significant difference between the lipoidemia and hemolysis groups in the measurement of HBV DNA RT-PCR (P>0.05). Conclusions Lipoidemia and hemolysis do not affect the results of HBV DNA RT-PCR.