Hepatitis B virus
Hepatitis B virus (HBV) affects millions of people worldwide and can lead to serious diseases such as liver cirrhosis and hepatocellular carcinoma. Highly sensitive detection of the viral DNA genome is possible with molecular methods. Genotyping of HBV is essential for characterization of patient groups and for epidemiological studies. There is increasing evidence that genotypic variation, mutations in the preCore region, and mutations associated with resistance to direct-acting antivirals are associated with efficacy of antiviral therapy and may require tailor-made treatment regimens. The polymerase chain reaction (PCR) in combination with advanced DNA sequencing technologies allows early and accurate detection of emerging resistant strains.
- Quantitative HBV-DNA (viral load)
- Quantitative HBV-RNA
- Amplification of HBV-DNA or HBV-RNA from plasma or liver samples (partial or full viral HBV genome)
- Sanger population sequencing (ABI), including data analysis and reporting
- Deep sequencing (Illumina MiSeq), including data analysis and reporting
- Reverse hybridization (LiPA)
- Based on PCR followed by direct sequence analysis
- Identification of all known genotypes (A-H)
- HBV genotyping can also be derived from HBV-RNA in cases where HBV-DNA is not detectable
Screening for mutations affecting the level of HBeAg expression
- Based on PCR followed by LiPA, or by direct sequence analysis
- Targeting mutations in the promoter region and at codon 28 of the preCore region
Screening for mutations associated with resistance to antiviral therapy
- Sanger or deep sequencing analysis of the HBV polymerase gene (full-length genome, or selected regions of interest, e.g., the 1032 bp RT-domain, including all amino acid positions associated with antiviral resistance).
- Monitoring of patients during antiviral therapy using population or deep sequencing.
- Determination of relatedness between different HBV isolates based on viral genome sequences
- For outbreak management and epidemiological studies
- Based on PCR followed by DNA sequencing analysis of the full genome or the 1032 bp polymerase RT-domain
- Serum or EDTA plasma. Heparinized blood can not be used.