Abstract: Objective: To evaluate the difference in the clinical performance of quantitating HBV DNA in the serum samples from the patients with chronic hepatitis B measured by the different commercial certified HBV DNA quantification kits.Methods:①The six mixed serum samples with HBV viral load were measured by five commercial certified kits(A, B, C, D and E), to compare the results. The six mixed serum samples and the two national HBV DNA serum standard materials(GBW-E-090137 and GBW-E-090139) were measured in three batches using the three domestic certified kits C, D and E, to verify the precision and accuracy of these three kits.②The serum samples from 43 patients with chronic hepatitis B were measured by the kits C, D and E, to compare the correlation and concordance of the results from these three different HBV DNA assays.Results:①Compared with A, there was no significant difference in the HBV DNA values of the six mixed serum samples measured by B or C(P>0.05). However, HBV DNA levels measured by D or E were significantly lower than the results by the kit A(P<0.05).②Resultsof the precision and accuracy tests showed that the coefficient of variation(CV) of the six mixed serum samples and the two national standard materials measured by the kits C, D, and E was less than 5%, and the differences between the expected and tested concentrations of these two national standard materials detected by the kits C, D, and E were all within 0.4 log₁₀ IU/mL, which showed high precision and accuracy.③The detection rates of the serum samples from 43 patients with chronic hepatitis B as determined by the kits C, D, and E were 95.35%(41/43), 95.35%(41/43), and 86.05%(37/43), respectively. Among the 43 clinical serum samples, 24 were quantifiable by three assays. For the 24 clinical serum samples, the quantitative results of the three assays were C>D>E(P<0.05); the correlation analysis indicated significant correlations between the three assays(C vs D: R²=0.93, Y=0.973 X-0.164; C vs E: R²=0.61, Y=0.770 X+0.210; D vs E: R²=0.69, Y=0.809 X+0.270); Bland-Altman plot analysis found mean differences of(C-D)=0.28 log₁₀ IU/mL(95% confidence interval,-0.30 to 0.86),(C-E)=0.80 log₁₀ IU/mL(95%CI,-0.62 to 2.21),(D-E)=0.51 log₁₀ IU/mL(95%CI,-0.74 to 1.77), respectively; the proportions of the specimens within the 95% limit of agreement were 95.83%(23/24) between C and D, 95.83%(23/24) between C and E, and 91.67%(22/24) between D and E, respectively. And the maximum differences within the 95% acceptable range were 0.85 log₁₀ IU/mL between C and D, 1.96 log₁₀ IU/mL between C and E, and 1.25 log₁₀ IU/mL between D and E, respectively. The proportions of the specimens with>1 log₁₀ IU/mL difference of HBV DNA levels were 0(0/24) between C and D, 33.33%(8/24) between C and E, and 25.00%(6/24) between D and E, respectively.Conclusion: There is a significant difference in the clinical performance of quantitating serum HBV DNA by the different commercial kits. All three HBV domestic certified assays have low commutability and may be not used interchangeably in routine clinical practice.