ArticleJournal of virological methods2025
High-dimensional droplet digital PCR of multiple hepatitis B splice variants in serum, liver, and tissue culture.
Article in Journal of virological methods, 2025. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 1 paper.
What it found
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The abstract states no effect estimate the extractor could read, or names no intervention and outcome on the map, so this paper lights no cell and moves no belief. It is still indexed, cited and linked below.
The trial behind it
Trials whose registry record cites this paper, or whose number appears in the abstract. A trial that started after this paper was published is citing it as background, not reporting it.
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Who cites it
1 citing paper in PubMed.
- Multi-Biofluid Approaches for cftDNA and cftRNA Biomarker Detection: Advances in Early Cancer Detection and Monitoring.Current issues in molecular biology · 2025Review
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Authors and funding
7 authors.
Funding
Abstract
Hepatitis B virus (HBV) affects approximately 2 billion individuals; 254 million have chronic infection. The roles of spliced HBV (spHBV) RNAs in human infection are poorly understood partly due to limitations in quantitative methods. We designed and multiplexed long amplicon (0.3-2kbp) ddPCR assays that simultaneously quantify the most frequently observed spHBV isoforms (sp1, sp2, sp3, sp8, sp9) alongside unspliced pregenomic RNA (pgRNA). Our innovations include the use of ddPCR, long amplicons, and high-dimensional multiplexing. We tested primers and probes using qPCR on synthetic DNA targets to identify a high-fidelity combination recognizing HBV genotype A viruses. Using common forward and reverse primers we accounted for PCR efficiency differences between amplicons of differing size. We altered denaturation and annealing temperatures to enrich spHBV, the minority population of total HBV RNA. To achieve higher-order multiplexing, we increased the number of cycles to 80, added a post-reverse transcription cleanup step, and tested > 360 oligo concentration combinations. We performed spatial transcriptomic sequencing on liver tissue to confirm splice junctions measured by ddPCR. We successfully quantified spHBV and pgRNA in serum, liver, and HBV-infected HepG2
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Registered trials
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