Expression the performance characteristics of TaqMan PCR assay for the detection and quantitation of HBV

Objective To characterize the performance characteristics of a quantitative real time TaqMan PCR assay,for the detection and quantitation of HBV genotypes A-G from patients' plasma and serum.This test was evaluated for limit of detection,dynamic range,reproducibility,accuracy,and genotype inclusivity. Methods Primers and TaqMan probes specific for HBV and an internal quantitation standard(QS)were bought from Louchi and tested using a set of plasmid DNAs representing various genotyped specimens.In addition,sensitivity,dynamic range,precision were evaluated using HBV dilution panels and patient specimens.Results and Conclusion A real time TaqMan assay was developed that detects and quantifies DNA from genotypes A-G equivalently.The assay has a limit of detection of 50 copies/ml with plasma and serum,a dynamic range up to 109 copies/ml,and good precision and accuracy.Titers obtained with this method without dilutions show good corelation across the dynamic range with titers obtained with this test.