Discussion of the performance verification procedures of the real-time quantitative polymerase chain reaction for determination of virus nucleic acid

Objective To discuss the reasonable performance verification procedures of real-time quantitative fluorescent polymerase chain reaction for determination of virus nucleic acid. Methods The performance verification procedures were applied according to the related documents published by Clinical and Laboratory Standards Institute(CLSI). The limits of quantification were verified by diluting the samples to which can not be tested. Results The within-run coefficient of variation(CV) of HBV-DNA were 4.28% and 2.08%, the total CV were 4.14% and 2.69% and the regression equation was y=1.1082x-0.5225; the correlation of linearity was 0.9976, the regression equation was y=0.9771x-0.0062, the linearity range was 1.05E3~1.05E8 and the limit of quantitation was 500 IU/mL. The within-run CV of HCV-RNA were 4.11% and 2.63%, the total CV were 5.15% and 3.41%, the regression equation was y=1.0075x-0.0662; the correlation of linearity was 0.9974,the regression equation was y=1.0479x-0.2594,the linearity range was 1.50E3~2.53E7 and the limit of quantitation was 1000 IU/mL. Conclusion It is necessary to run the performance verification procedures before using new tests in clinical laboratory. The performance verification procedures of quantitative PCR for testing nucleic virus and the statistics methods are selected on the basis of practical situations.