HBV whole-genome sequencing using tiling PCR and nanopore sequencing
We evaluated an adapted tiling PCR approach for near-complete hepatitis B virus (HBV) genome sequencing from clinical serum samples using Oxford Nanopore long-read sequencing. A previously published primer scheme (Ringlander et al. 2022), originally developed for Ion Torrent sequencing, was modified by removing platform-specific adapter sequences and applied to nanopore sequencing. Clinical serum samples from patients with chronic HBV infection were processed. Full-genome amplification was performed using overlapping amplicons spanning the ~3.2 kb HBV genome, followed by sequencing on a GridION instrument using the Rapid Sequencing Kit. Sequencing reads were processed using the ARTIC bioinformatics pipeline for primer trimming, read mapping, and consensus genome generation. Samples representing HBV genotypes A–E were included to capture genetic diversity. This study evaluates the performance of an adapted tiling PCR scheme across a range of viral concentrations and genotypes. Human reads were removed prior to submission.
We evaluated an adapted tiling PCR approach for near-complete hepatitis B virus (HBV) genome sequencing from clinical serum samples using Oxford Nanopore long-read sequencing. A previously published primer scheme (Ringlander et al. 2022), originally developed for Ion Torrent sequencing, was modified by removing platform-specific adapter sequences and applied to nanopore sequencing. Clinical serum samples from patients with chronic HBV infection were processed. Full-genome amplification was performed using overlapping amplicons spanning the ~3.2 kb HBV genome, followed by sequencing on a GridION instrument using the Rapid Sequencing Kit. Sequencing reads were processed using the ARTIC bioinformatics pipeline for primer trimming, read mapping, and consensus genome generation. Samples representing HBV genotypes A–E were included to capture genetic diversity. This study evaluates the performance of an adapted tiling PCR scheme across a range of viral concentrations and genotypes. Human reads were removed prior to submission.
ELIXIR Norway