To evaluate the measurement uncertainty (MU) of real-time fluorescent quantitative PCR detection of HBV DNA and HCV RNA according to laboratory accreditation requirements of ISO15189.

The MU introduced by the reproducibility of HBV DNA and HCV RNA measurements was calculated using internal quality control. The MU introduced by the bias of HBV DNA and HCV RNA was calculated by using the proficiency testing results of National Center for Clinical Laboratories.

The relative synthesis standard uncertainty for HBV DNA determination at low concentrations was 5.21%, while at high concentrations it was 3.47%. The relative expanded uncertainty at low concentrations was 10.42%, while at high concentrations it was 6.94% (k=2). The relative synthesis standard uncertainty for HCV RNA determination at low concentrations was 6.43%, and at high concentrations it was 5.18%. The relative expanded uncertainty at low concentrations was 12.86%, while at high concentrations it was 10.36% (k=2). The expanded uncertainty of HBV DNA at low concentration (=3.45) was 0.36, which was less than the target uncertainty (0.40) , but at high concentration (=6.38) the expansion uncertainty was 0.44, higher than the target uncertainty (0.40). The expansion uncertainty of HCV RNA at low (=3.23) and high (=5.24) concentrations was 0.42 and 0.54, respectively, both higher than the target uncertainty (0.40).

The MU introduced by real-time fluorescent quantitative PCR determination of HBV DNA and HCV RNA makes the results comparable, which is helpful to the interpretation of the clinical results and can be used as the basis for the continuous improvement of the quality.