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Updated: Aug 20, 2026
Real-Time Polymerase Chain Reaction-Based Detection and Quantification of Hepatitis B Virus DNA
Published on: December 15, 2023
Detection of hepatitis B virus DNA by real-time PCR using TaqMan-MGB probe technology
Jin-Rong Zhao1, Yu-Jie Bai, Qing-Hua Zhang
1Institute of Genetic Diagnosis, Fourth Military Medical University, 17 Changle West Road, Xi'an 710032, Shaanxi Province, China. zhaojrr@fmmu.edu.cn
Aim:
To develop a real-time PCR for detecting hepatitis B virus (HBV) DNA based on TaqMan technology using a new MGB probe.
Methods:
Plasmid containing the sequence of X gene (1414-1744 nt) was constructed as HBV-DNA standard for quantitative analysis. A TaqMan-MGB probe between primers for amplification was designed to detect PCR products. The interested sequence contained in the plasmid and in clinical specimens was quantitatively measured.
Results:
The detection limit of the assay for HBV DNA was 1 genome equivalent per reaction. A linear standard curve was obtained between 10(0) and 10(9) DNA copies/reaction (r>0.990). None of the negative control samples showed false-positive reactions in duplicate. HBV DNA was detected in 100% (50/50) of HBV patients with HbeAg, and in 72.0% (36/50) with HBsAg, HBeAb and HBcAb. The coefficient of variation for both intra- and inter-experimental variability demonstrated high reproducibility and accuracy.
Conclusion:
Real-time PCR based on TaqMan-MGB probe technology is an excellent method for detection of HBV DNA.
