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<p>The detection of circulating tumor <i>DNA</i> (<i>ctDNA</i>) with high sensitivity and specificity is crucial for the early diagnosis and monitoring of tumors, as well as for drug therapy. In this study, a simple and highly sensitive biosensor was specifically designed for the identification of targeted <i>ctDNA</i>. For the first time, a three-dimensional polyvinylidene fluoride-graphene oxide-chitosan (PVDF/CS/GO) nanofiber mesh was fabricated on a polydimethylsiloxane (PDMS) micropillar substrate using electrospinning technology, and the nanofibers were functionalized with peptide nucleic acids probe-gold nanoparticle (<i>PNA</i>-AuNP) complexes, which served as affinity molecules for detecting the methylation of the <i>E542K</i> variant of the phosphatidylinositol-4,5-bisphosphate 3-kinase catalytic subunit α (<i>PIK3CA</i>) gene in the peripheral blood of cancer patients. Additionally, an anti-5-Methylcytosine monoclonal antibody - multi-walled carbon nanotubes-COOH complex (Anti-5-mC-MWCNTs-COOH) complex was incubated to result in significantly amplified electrochemical signals for the accurate quantification of the <i>E542K</i> variant of the <i>PIK3CA</i> gene. Detectable signal responses were observed only when both molecules were simultaneously present, greatly enhancing the accuracy of the analysis. The biosensor exhibits high capture sensitivity for the methylation level of the <i>E542K</i> variant of the <i>PIK3CA</i> gene across a concentration range of 50 to 10 000 fmol/L, with the lowest detection limit of 10 fmol/L. The <i>ctDNA</i> nanobiosensor has been shown to be both feasible and valuable for quantifying <i>ctDNA</i> concentrations in clinical blood samples. Consequently, this 3D nanofiber biosensor shows significant potential for clinical applications in cancer diagnosis and personalized medical treatments.</p>

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A Novel Electrochemical Biosensor: Sensitive and Quantitative Detection of Circulating Tumor DNA

  • 期刊论文

  • 发表日期：

    2025年10月29日

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  • Yujuan Wu,
  • Yu Li,
  • Jianyu Mao,
  • Weiying Zhang,
  • Qinqin Ran,
  • Yali Ben

摘要

The detection of circulating tumor DNA (ctDNA) with high sensitivity and specificity is crucial for the early diagnosis and monitoring of tumors, as well as for drug therapy. In this study, a simple and highly sensitive biosensor was specifically designed for the identification of targeted ctDNA. For the first time, a three-dimensional polyvinylidene fluoride-graphene oxide-chitosan (PVDF/CS/GO) nanofiber mesh was fabricated on a polydimethylsiloxane (PDMS) micropillar substrate using electrospinning technology, and the nanofibers were functionalized with peptide nucleic acids probe-gold nanoparticle (PNA-AuNP) complexes, which served as affinity molecules for detecting the methylation of the E542K variant of the phosphatidylinositol-4,5-bisphosphate 3-kinase catalytic subunit α (PIK3CA) gene in the peripheral blood of cancer patients. Additionally, an anti-5-Methylcytosine monoclonal antibody - multi-walled carbon nanotubes-COOH complex (Anti-5-mC-MWCNTs-COOH) complex was incubated to result in significantly amplified electrochemical signals for the accurate quantification of the E542K variant of the PIK3CA gene. Detectable signal responses were observed only when both molecules were simultaneously present, greatly enhancing the accuracy of the analysis. The biosensor exhibits high capture sensitivity for the methylation level of the E542K variant of the PIK3CA gene across a concentration range of 50 to 10 000 fmol/L, with the lowest detection limit of 10 fmol/L. The ctDNA nanobiosensor has been shown to be both feasible and valuable for quantifying ctDNA concentrations in clinical blood samples. Consequently, this 3D nanofiber biosensor shows significant potential for clinical applications in cancer diagnosis and personalized medical treatments.