4.7 Article

Improved hepatitis delta virus genome characterization by single molecule full-length genome sequencing combined with VIRiONT pipeline

期刊

JOURNAL OF MEDICAL VIROLOGY
卷 95, 期 3, 页码 -

出版社

WILEY
DOI: 10.1002/jmv.28634

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genotyping; HDV editing; hepatitis delta virus (HDV); local pipeline; long-read sequencing; whole-genome sequencing

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This study presents a workflow to amplify, sequence, and analyze the whole HDV genome in a single fragment. The HDV genome was successfully amplified and fully sequenced, allowing accurate subtyping and identification of a new HDV genotype 1 subtype. This workflow overcomes genome assembly issues and helps to identify modifications throughout the whole HDV genome.
Hepatitis B virus (HBV) and hepatitis D virus (HDV) coinfection confers a greater risk for accelerated liver disease progression. Full-length characterization of HDV genome is necessary to understand pathogenesis and treatment response. However, owing to its high variability and tight structure, sequencing approaches remain challenging. Herein, we present a workflow to amplify, sequence, and analyze the whole HDV genome in a single fragment. Sequencing was based on the Oxford Nanopore Technologies long-read sequencing followed by a turnkey analysis pipeline (VIRiONT, VIRal in-house ONT sequencing analysis pipeline) that we developed and make available online for free. For the first time, HDV genome was successfully amplified and full-length sequenced in a single fragment, allowing accurate subtyping from 30 clinical samples. High variability of edition, a crucial step in viral life cycle, was found among samples (from 0% to 59%). Additionally, a new subtype of HDV genotype 1 was identified. We provide a complete workflow for assessment of HDV genome at full-length quasispecies resolution overcoming genome assembly issues and helping to identify modifications throughout the whole genome. This will help a better understanding of the impact of genotype/subtype, viral dynamics, and structural variants on HDV pathogenesis and treatment response.

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