Evaluation of a novel domestic hepatitis B viral DNA quantitative fluorescence diagnostic kit

Objective Comparative performance characteristics of a novel domestic (Hunan Sansure) HBV DNA quantitative fluorescence diagnostic kit (PCR-fluorescence probing) and its counterpart Roche Cobas Ampliprep/COBAS Taqman HBV test kit. Methods Firstly,WHO international standard for HBV-DNA was diluted to 50,200,2000,20 000 IU/ml separately,and then all diluted samples were detected by both the novel domestic HBV DNA quantitative fluorescence diagnostic kit and Roche Cobas Ampliprep/COBAS Taqman HBV test kit to perform traceability analysis and to determine kits′reliability. Secondly,pre-diluted series of standard HBV B and C genotype plasma (25, 50, 200, 2000, 20 000, 200 000 IU/ml) were simultaneously performed 216 times for three-center detection by use of the domestic and Roche HBV DNA test kits separately.Accuracy and precision of those two types of HBV DNA kits were comparatively analyzed.Finally,a total of 42 clinical plasma samples including 5 negative HBV DNA and different concentration levels of 37 positive HBV DNA were detected by the above two types of kits to perform clinical quality evaluation. Results Traceability results showed that both HBV DNA test kits agreed with the HBV DNA WHO standard across all four titers tested and all absolute differences (observed mean HBV HBV-expected HBV DNA) were within 0.3 lg IU/ml.Accuracy results indicated that thedeviation of all observed values at 6 titers (both of HBV genotyping B and C) tested were within the acceptance criteria (0.005-0.280 lg IU/ml).Comparative performance of coefficient of variation of PCR assays to HBV genotypes B and C measured by both kits showed that there were no differences of precision were found (genotyping B: t=1.226,P=0.275; genotyping C: t=2.319,P=0.07).The clinical performance of the domestic assay compared to the COBAS assay had been assessed on a panel of 42 clinical specimens.The qualitative results indicated that the total concordant results between two tests were determined in 97.62% (41/42) of samples.Also,a significant correlation was observed between values tested by two HBV DNA kits in 37 HBV DNA positive samples (R2=0.934,P<0.0001). Conclusion No significant differences of the traceability,accuracy,precision and clinical detection are found between two kits.(Chin J Lab Med,2013,36:280-285) Key words: Hepatitis B virus; DNA; viral; Reagent kits; diagnostic; Eevaluation studies