Related Experiment Video
Updated: Jun 13, 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 genotypes A to D by the fluorescence polarization assay based on asymmetric PCR
Li Ding1, Chen Rui, Jia Zhansheng
1The Center of Biotechnological Diagnosis and Therapy, The 261st Hospital of PLA, Beijing, China.
Insights
A new, cost-effective fluorescence polarization method accurately detects hepatitis B virus (HBV) genotypes A to D. This simplifies HBV genotyping, overcoming technical challenges of current methods.
Area of Science:
- Virology
- Molecular Biology
- Diagnostic Assays
Background:
- Hepatitis B virus (HBV) genotypes A and D are globally prevalent, while B and C dominate in Asia.
- HBV genotypes C and D are associated with poorer treatment response compared to A and B.
- Existing HBV genotype detection methods (A-D) are often costly and technically demanding for routine use.
Purpose of the Study:
- To develop a simple, cost-effective method for simultaneous detection of hepatitis B virus genotypes A to D.
- To establish a novel fluorescence polarization (FP) assay for HBV genotyping.
Main Methods:
- Utilized asymmetric PCR with general primers targeting the preS region of HBV.
- Employed four distinct fluorophore-labeled probes specific for HBV genotypes A-D.
- Measured genotype-specific hybridization by changes in fluorescence polarization (FP) values.
Main Results:
- The developed FP assay successfully differentiated between HBV genotypes A, B, C, and D.
- A parallel study comparing FP assay with type-specific PCR on 1398 samples showed no significant difference in results.
- The FP method demonstrated high accuracy and reliability for HBV genotyping.
Conclusions:
- A simple, cost-effective fluorescence polarization assay for simultaneous detection of HBV genotypes A-D has been successfully developed.
- This novel method offers a viable alternative to conventional techniques, overcoming technical and cost barriers.
- The FP assay provides accurate and reliable HBV genotyping, facilitating wider clinical application and research.
Abstract:
The hepatitis B virus genotypes A and D have global distributions, while the hepatitis B genotypes B and C are predominant in Asia. Individuals infected with genotype C or D have a lower response rate to therapy than individuals infected with genotype A or B. The conventional detections of hepatitis B genotypes A to D, however, have not been used routinely due to technical difficulties and high costs. A simple and cost-effective method for simultaneous detection of the hepatitis B genotypes A to D has been developed. A pair of general primers in the preS region of the hepatitis B were used in an asymmetric PCR. Four probes specific for the hepatitis B genotypes A to D labeled with different fluorophores hybridized, respectively with their target amplicons, and the hybridization increased the fluorescence polarization (FP) values. The genotypes were determined by the increased fluorescence polarization values. DNA extracted from 1398 samples was detected by the FP and the type-specific PCR assay in parallel. No significance difference was found between the two methods.
