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Background <p>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.</p> Methods <p>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.</p> Results <p>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.</p> Conclusions <p>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 dynamics and supporting personalized treatment strategies.</p>

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

  • 期刊论文

  • 出版模式：

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  • 发表日期：

    2025年10月10日

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  • Jingmei Li,
  • Hui Li,
  • Xin Zhang,
  • Xueyan Liu,
  • Zumi Zhou,
  • Jia Wei,
  • Rujuan Zhao,
  • Tai-Cheng Zhou,
  • Zijie Zhang,
  • Chunmei Li

摘要

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 dynamics and supporting personalized treatment strategies.