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<p>Tumor cell subtyping is clinically critical for early cancer diagnosis and prognostic assessment. As key regulators of cellular phenotype, microRNAs (miRNAs) exhibit cell-specific expression profiles that serve as pivotal tumor classification biomarkers. However, current multiplex miRNA detection technologies suffer from notable limitations. Quantitative polymerase chain reaction (qPCR) is vulnerable to primer cross-amplification interference, and sequencing and microdevice-based methods are either cost-prohibitive or operationally cumbersome. These drawbacks severely limit their clinical applicability for rapid, user-friendly diagnostics. To address these limitations, this study proposes a novel DNA fiber barcode-based multiplex miRNA detection strategy for accurate tumor cell subtyping. Programmable DNA tile self-assembly constructs functional DNA fibers with target-specific recognition units. Binding of miRNAs triggers conformational changes in the hairpin probe, generating detectable fluorescent barcode signals that enable the simultaneous identification and quantification of miRNAs. Experiments demonstrate a robust linear response within a range of 10 pM–70 nM, with detection limits between 2.4 pM and 5.0 pM for the analysis of miR-21, miR-105, and miR-155. The results align consistently with qPCR findings for cancer and normal cell lines, confirming the method’s accuracy and reliability. This strategy overcomes existing technical limitations, provides new insights for miRNA-based cellular analysis, and establishes both theoretical and technical foundations for precision tumor subtyping.</p>

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Multiple miRNA Analysis Based on DNA Fiber Barcodes for Cell Typing

  • 期刊论文

  • 发表日期：

    2026年5月20日

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  • Xiaoxing Fang,
  • Xuewei Li,
  • Wenqi Gao,
  • Chao Li,
  • Ji Zheng

摘要

Tumor cell subtyping is clinically critical for early cancer diagnosis and prognostic assessment. As key regulators of cellular phenotype, microRNAs (miRNAs) exhibit cell-specific expression profiles that serve as pivotal tumor classification biomarkers. However, current multiplex miRNA detection technologies suffer from notable limitations. Quantitative polymerase chain reaction (qPCR) is vulnerable to primer cross-amplification interference, and sequencing and microdevice-based methods are either cost-prohibitive or operationally cumbersome. These drawbacks severely limit their clinical applicability for rapid, user-friendly diagnostics. To address these limitations, this study proposes a novel DNA fiber barcode-based multiplex miRNA detection strategy for accurate tumor cell subtyping. Programmable DNA tile self-assembly constructs functional DNA fibers with target-specific recognition units. Binding of miRNAs triggers conformational changes in the hairpin probe, generating detectable fluorescent barcode signals that enable the simultaneous identification and quantification of miRNAs. Experiments demonstrate a robust linear response within a range of 10 pM–70 nM, with detection limits between 2.4 pM and 5.0 pM for the analysis of miR-21, miR-105, and miR-155. The results align consistently with qPCR findings for cancer and normal cell lines, confirming the method’s accuracy and reliability. This strategy overcomes existing technical limitations, provides new insights for miRNA-based cellular analysis, and establishes both theoretical and technical foundations for precision tumor subtyping.