Related Experiment Video
Updated: Jul 19, 2026
Real-Time Polymerase Chain Reaction-Based Detection and Quantification of Hepatitis B Virus DNA
Published on: December 15, 2023
Simultaneous quantification and genotyping of hepatitis B virus for genotypes A to G by real-time PCR and two-step
Wen-Chun Liu1, Masashi Mizokami, Maria Buti
1Institute of Basic Medical Sciences, Department of Medicine, National Cheng Kung University, Tainan City 704, Taiwan, Republic of China, and Liver Unit, Hospital General Universitari Vall d'Hebron, Barcelona, Spain.
Abstract:
Both the viral titer and the genotype significantly determine clinical outcomes and responses to antiviral treatment in chronic hepatitis B virus (HBV) infection. A method was developed for large-scale A-to-G genotyping with simultaneous viral quantification. The assay was run on a LightCycler instrument using hybridization probes. The genotype was determined from the melting points of the probes in a two-step manner. Set 1 amplicons differentiated genotypes B, E, and F from A, C, D, and G and simultaneously quantified viremia by real-time PCR. Melting curve analysis using the set 2-1 amplicon or the set 2-2 amplicon reaction mixture was then used to differentiate these genotype groups into single genotypes. HBV DNA quantification was consistent with that of the Amplicor assay and linear in a range from 10(2) to 10(13) copies/ml. By comparison with the restriction fragment length polymorphism method, 92.3% of 441 samples were accurately genotyped by the current assay. The method should be useful for genotyping and quantification of HBV DNA in areas where all genotypes exist.
