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Abstract <p><b>Objective:</b> Develop a PCR-free DNA nanomachine (BDM) for selective picomolar detection of plasma miR-371a. <b>Methods:</b> A split 10–23 deoxyribozyme was engineered with substrate-delivering hooks and mismatch-introducing arms. The system was validated at 35°C using synthetic targets and patient plasma. <b>Results and Discussion:</b> BDM achieved a 1.6 pM limit of detection, showing a 9-fold sensitivity increase over hook-less controls. It maintained excellent specificity (LOD 1.3 pM) against a 25-fold excess of homologs and successfully identified clinical seminoma samples. <b>Conclusions:</b> BDM offers a sensitive, contamination-resistant alternative to RT-qPCR for non-invasive cancer liquid biopsies.</p>

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A Binary DNAzyme Machine (BDM) for Selective Amplification-Free Detection of microRNA

  • 期刊论文

  • 发表日期：

    2026年7月16日

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  • Meera Alaji,
  • Ahmed A. Eldeeb,
  • A. Shalaev,
  • A. Malek,
  • Moustapha A. Y. Nour

摘要

Abstract

Objective: Develop a PCR-free DNA nanomachine (BDM) for selective picomolar detection of plasma miR-371a. Methods: A split 10–23 deoxyribozyme was engineered with substrate-delivering hooks and mismatch-introducing arms. The system was validated at 35°C using synthetic targets and patient plasma. Results and Discussion: BDM achieved a 1.6 pM limit of detection, showing a 9-fold sensitivity increase over hook-less controls. It maintained excellent specificity (LOD 1.3 pM) against a 25-fold excess of homologs and successfully identified clinical seminoma samples. Conclusions: BDM offers a sensitive, contamination-resistant alternative to RT-qPCR for non-invasive cancer liquid biopsies.