Advanced molecular surveillance approaches for characterization of blood borne hepatitis viruses

PLoS One. 2020 Jul 17;15(7):e0236046. doi: 10.1371/journal.pone.0236046. eCollection 2020.

Abstract

Defining genetic diversity of viral infections directly from patient specimens is the ultimate goal of surveillance. Simple tools that can provide full-length sequence information on blood borne viral hepatitis viruses: hepatitis C, hepatitis B and hepatitis D viruses (HCV, HBV and HDV) remain elusive. Here, an unbiased metagenomic next generation sequencing approach (mNGS) was used for molecular characterization of HCV infections (n = 99) from Israel which yielded full-length HCV sequences in 89% of samples, with 7 partial sequences sufficient for classification. HCV genotypes were primarily 1b (68%) and 1a (19%), with minor representation of genotypes 2c (1%) and 3a (8%). HBV/HDV coinfections were characterized by suppressed HBV viral loads, resulting in sparse mNGS coverage. A probe-based enrichment approach (xGen) aiming to increase HBV and HDV coverage was validated on a panel of diverse genotypes, geography and titers. The method extended HBV genome coverage a median 61% (range 8-84%) and provided orders of magnitude boosts in reads and sequence depth for both viruses. When HBV-xGen was applied to Israeli samples, coverage was improved by 28-73% in 4 samples and identified HBV genotype A1, A2, D1 specimens and a dual B/D infection. Abundant HDV reads in mNGS libraries yielded 18/26 (69%) full genomes and 8 partial sequences, with HDV-xGen only providing minimal extension (3-11%) of what were all genotype 1 genomes. Advanced molecular approaches coupled to virus-specific capture probes promise to enhance surveillance of viral infections and aid in monitoring the spread of local subtypes.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Blood / virology*
  • Cohort Studies
  • Genotype
  • Hepatitis Viruses / genetics*
  • Hepatitis Viruses / isolation & purification
  • High-Throughput Nucleotide Sequencing*
  • Humans
  • Metagenomics*

Grants and funding

The authors declare we were funded by Abbott Laboratories and that one or more of the authors have an affiliation to the commercial funders of this research study, Abbott Laboratories. The funder provided support in the form of salaries for authors [MGB, AO, KF, BJH, JY, MAR, GAC], but did not have any additional role in the study design, data collection and analysis, decision to publish, or preparation of the manuscript. The specific roles of these authors are articulated in the ‘author contributions’ section.