ArticleJournal of virology2026
A novel highly specific qPCR assay for distinguishing HBV cccDNA from rcDNA following bisulfite pretreatment.
Article in Journal of virology, 2026. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Not yet cited in PubMed.
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16 authors.
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Abstract
The persistence of covalently closed circular DNA (cccDNA) underlies chronic hepatitis B virus (HBV) infection and remains as the major barrier to achieving a functional cure. An accurate and sensitive quantification of cccDNA is critical for evaluating therapeutic strategies. The conventional quantitative PCR (qPCR) lacks specificity to distinguish cccDNA from viral relaxed circular DNA (rcDNA) and other replicative intermediates, while nuclease pretreatment (e.g., plasmid-safe ATP-dependent nuclease, exonucleases I/III, or T5 exonuclease) may either incompletely digest rcDNA and/or risk over-digesting cccDNA. Here, we evaluated a novel patented bisulfite conversion-based cccDNA (BSC-cccDNA) qPCR platform that employs bisulfite treatment to disrupt rcDNA complementarity, followed by qPCR with primer sets targeting the rcDNA gap region, to enable selective amplification of cccDNA. Using synthetic HBV DNA controls and DNA samples extracted from HBV stable cell line and HBV-infected cells, we demonstrate that the BSC-cccDNA qPCR assay achieves high specificity, with tolerance of up to 10
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