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<p>Alterations in microRNA (miRNA) expression profiles play a pivotal role in the initiation and progression of various diseases, including pediatric pneumonia. Consequently, the development of sensitive, specific, and precise methodologies for miRNA detection represents a promising tool for clinical diagnosis and therapeutic intervention. This study proposes a highly sensitive miRNA detection approach utilizing catalytic hairpin assembly (CHA)-mediated probe release through chain extension and cascade signal amplification. The method enables precise discrimination of target miRNAs from other RNA species via a dual-target recognition mechanism. Furthermore, the incorporation of four signal amplification cycles confers substantial amplification efficiency, permitting accurate detection of low-abundance miRNAs. This system employs cascade isothermal amplification, demonstrating a linear response across concentrations from 1 fM to 100&#xa0;pM and achieving a detection limit of 0.54 fM. The methodology exhibits significant potential for practical application in complex biological samples, providing a novel platform for profiling miRNA expression patterns and elucidating their mechanistic roles in disease pathogenesis.</p>

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Loading scaffold-mediated multiple signal cycle for ultrasensitive microRNA analysis with low background signal

  • 期刊论文

  • 出版模式：

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  • 发表日期：

    2025年8月12日

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  • Zhe Zhang,
  • Qiaoyi Xie,
  • Chenbo Zhu,
  • Huali Shao,
  • Boying Wu

摘要

Alterations in microRNA (miRNA) expression profiles play a pivotal role in the initiation and progression of various diseases, including pediatric pneumonia. Consequently, the development of sensitive, specific, and precise methodologies for miRNA detection represents a promising tool for clinical diagnosis and therapeutic intervention. This study proposes a highly sensitive miRNA detection approach utilizing catalytic hairpin assembly (CHA)-mediated probe release through chain extension and cascade signal amplification. The method enables precise discrimination of target miRNAs from other RNA species via a dual-target recognition mechanism. Furthermore, the incorporation of four signal amplification cycles confers substantial amplification efficiency, permitting accurate detection of low-abundance miRNAs. This system employs cascade isothermal amplification, demonstrating a linear response across concentrations from 1 fM to 100 pM and achieving a detection limit of 0.54 fM. The methodology exhibits significant potential for practical application in complex biological samples, providing a novel platform for profiling miRNA expression patterns and elucidating their mechanistic roles in disease pathogenesis.