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High serum levels of pregenomic RNA reflect frequently failing reverse transcription in hepatitis B virus particles

Journal article
Authors Kasthuri Prakash
Gustaf E Rydell
Simon B. Larsson
Maria Andersson
Gunnar Norkrans
Helene Norder
Magnus Lindh
Published in Virology Journal
Volume 15
ISSN 1743-422X
Publication year 2018
Published at Institute of Biomedicine, Department of Infectious Medicine
Language en
Keywords surface-antigen, hepatocellular-carcinoma, mutations, carriers, disease, hbsag
Subject categories Microbiology in the medical area


Background: Hepatocytes infected by hepatitis B virus (HBV) produce different HBV RNA species, including pregenomic RNA (pgRNA), which is reverse transcribed during replication. Particles containing HBV RNA are present in serum of infected individuals, and quantification of this HBV RNA could be clinically useful. Methods: In a retrospective study of 95 patients with chronic HBV infection, we characterised HBV RNA in serum in terms of concentration, particle association and sequence. HBV RNA was detected by real-time PCR at levels almost as high as HBV DNA. Results: The HBV RNA was protected from RNase and it was found in particles of similar density as particles containing HBV DNA after fractionation on a Nycodenz gradient. Sequencing the epsilon region of the RNA did not reveal mutations that would preclude its binding to the viral polymerase before encapsidation. Specific quantification of precore RNA and pgRNA by digital PCR showed almost seven times lower ratio of precore RNA/pgRNA in serum than in liver tissue, which corresponds to poorer encapsidation of this RNA as compared with pgRNA. The serum ratio between HBV DNA and HBV RNA was higher in genotype D as compared with other genotypes. Conclusions: The results suggest that HBV RNA in serum is present in viral particles with failing reverse transcription activity, which are produced at almost as high rates as viral particles containing DNA. The results encourage further studies of the mechanisms by which these particles are produced, the impact of genotype, and the potential clinical utility of quantifying HBV RNA in serum.

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