A single-cell digital PCR method tailored for quantification of HBV DNA positive cells in liver biopsy tissues

作者：Jingmei Li, Hui Li, Xin Zhang, Xueyan Liu, Zumi Zhou, Jia Wei, Rujuan Zhao, Taicheng Zhou, Zijie Zhang, Chunmei Li · 发表于：Journal of Translational Medicine · 年份：2025 · DOI：10.1186/s12967-025-07131-9 · 被引用次数：3 · 研究领域：Hepatitis B Virus Studies、Hepatitis C virus research、Innovative Microfluidic and Catalytic Techniques Innovation

BACKGROUND: Accurate quantification of hepatitis B virus (HBV) DNA in hepatocytes is essential for understanding the virus's biology and improving treatment strategies. Existing methods lack the resolution to assess infection heterogeneity at the single-cell level, which is critical for evaluating treatment efficacy and guiding personalized therapy. METHODS: We developed a single-cell infection detection PCR (scID-PCR) method for quantifying intrahepatic HBV-positive cells with single-cell resolution. scID-PCR was validated using HepG2.2.15 and Hep3B2.1-7 cell models to assess sensitivity, specificity, and reproducibility. As proof of concept, scID-PCR was applied to liver biopsy samples from five chronic hepatitis B (CHB) patients and one non-HBV-related hepatocellular carcinoma (HCC) patient. RESULTS: Validation demonstrated that scID-PCR was highly sensitive, specific, and reproducible. scID-PCR detected HBV-positive cells in all CHB patients, including those with undetectable serum HBV DNA levels after 48 weeks of antiviral therapy. No cccDNA-positive cells were identified in the HCC patient. The proportion of HBV-positive cells offering insights into infection status and identifying patients more likely to achieve a cure. CONCLUSIONS: The scID-PCR platform offers a novel and powerful approach to quantify intrahepatic HBV-positive cells with single-cell resolution. This method complements traditional serological markers offering a more nuanced view of CHB infection dynami...