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Circulating Hepatitis B Virus Nucleic Acids in Chronic Infection

Representation of Differently Polyadenylated Viral Transcripts during Progression to Nonreplicative Stages

Department of Virus-Host Interactions [Q. S., R. B., C. H. S.] and Biostatistics Unit [L. E.], Deutsches Krebsforschungszentrum, 69120 Heidelberg, Germany; Departments of Pathology [Q. S., S-F. W.] and Clinical Diagnostics [T-E. C.], Tangdu Hospital, The Fourth Military Medical University, Xi’an, 710038 China; University of Lübeck, 23538 Lübeck, Germany [H. H.]; and Takegoshi Internal Medicine Clinic, Takaoka, 9330014 Japan [K. T.]

Purpose: Beside the established maturation of hepatitis B virus (HBV) transcripts at a polyadenylation signal downstream of the HBV x protein open reading frame, maturation at an internal polyadenylation signal has been observed in the chronically infected liver. In the present study, it was the aim to identify the respective circulating full-length and truncated transcripts in plasma/serum of carriers.

Experimental Design: Nucleic acids extracted from sera were analyzed using established PCR and reverse transcription-PCR procedures targeted to HBV x protein gene regions. Amplification products were cloned and sequenced.

Results: Base substitution patterns were determined, which indicated infection stages advanced to different degrees regardless of the transcript type analyzed. HBV full-length RNA (fRNA) showed a high correlation with hepatitis B e antigen and viral DNA, indicative for a replicative infection. In contrast, truncated RNA (trRNA) appeared to be independent of hepatitis B e antigen and showed only a weak association with circulating viral DNA. No correlation was observed between the levels of trRNA and the apparent liver damage as reflected by alanine transaminase levels. An age-dependent representation of fRNA and trRNA was observed: fRNA decreased progressively to low levels, whereas trRNA remained at comparably high values. trRNA and RNA not polyadenylated at either of the two polyadenylation signals were detected even in the absence of any other conventional HBV seromarker, including viral DNA. This was shown for patients with cryptogenic cirrhosis and hepatitis C virus carriers.

Conclusions: The identification of HBV RNA in human serum has a diagnostic potential for apparent and for inapparent infection stages.

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