Deep sequencing of hepatitis B virus (HBV) genomes using rolling circle amplification and Nanopore

The optimisation of unbiased deep-sequencing methods for full-length HBV genomes will inform detailed insights into the evolution and diversity of this pathogen. Nanopore-based sequencing technologies offer the potential for real-time detection and sequencing of blood-borne viruses, useful for clinical diagnosis, drug-resistance calling and high-resolution molecular epidemiology. To date, sequencing accuracy has limited the full potential of Oxford Nanopore’s long-read capabilities. We here describe use of an isothermal rolling-circle amplification (RCA) method, (i) to amplify HBV DNA and (ii) to derive concatemers of whole HBV genomes thus providing a mechanism to correct sequencing errors.