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Robust nontarget DNA-triggered catalytic hairpin assembly amplification strategy for the improved sensing of microRNA in complex biological matrices
Analyst ( IF 3.6 ) Pub Date : 2023-10-17 , DOI: 10.1039/d3an01411h Ruining Yang 1 , Xingfen Liu 1 , Junbo Hu 1 , Hui Xu 2 , Jixiang Song 1 , Huiyu Zhou 1 , Meixing Li 1 , Yanqin Huang 1 , Lei Zhang 1 , Quli Fan 1
Analyst ( IF 3.6 ) Pub Date : 2023-10-17 , DOI: 10.1039/d3an01411h Ruining Yang 1 , Xingfen Liu 1 , Junbo Hu 1 , Hui Xu 2 , Jixiang Song 1 , Huiyu Zhou 1 , Meixing Li 1 , Yanqin Huang 1 , Lei Zhang 1 , Quli Fan 1
Affiliation
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A simple but robust fluorescence strategy based on a nontarget DNA-triggered catalytic hairpin assembly (CHA) was constructed to probe microRNA-21 (miR-21). A short ssDNA rather than degradable target miRNA was employed as an initiator. Two molecular beacons needed to assist the CHA process were simplified to avoid unfavorable nonspecific interactions. In the presence of the target, the initiator was released from a partially duplex and triggered the cyclic CHA reaction, resulting in a significantly amplified optical readout. A wide linear range from 0.1 pM to 1000 pM for the sensing of miR-21 in buffer was achieved with a low detection limit of 0.76 pM. Fortunately, this strategy demonstrated an obviously improved performance for miR-21 detection in diluted serum. The fluorescence signals were enhanced remarkably and the sensitivity was further improved to 0.12 pM in 10% serum. The stability for miR-21 quantification and the capability for the analysis of single nucleotide polymorphisms (SNPs) were also improved greatly. More importantly, the biosensor could be applied to image miR-21 in different living tumor cells with high resolution, illustrating its promising potential for the assay of miRNAs in various complex situations for early-stage disease diagnosis and biological studies in cells.
中文翻译：
稳健的非靶标 DNA 触发催化发夹组装扩增策略，用于改善复杂生物基质中 microRNA 的传感
构建了一种基于非靶标 DNA 触发的催化发夹组装 (CHA) 的简单但稳健的荧光策略来探测 microRNA-21 (miR-21)。使用短 ssDNA 作为引发剂，而不是可降解的靶 miRNA。简化了辅助 CHA 过程所需的两个分子信标，以避免不利的非特异性相互作用。在靶标存在的情况下，引发剂从部分双链体中释放出来并触发循环 CHA 反应，从而产生显着放大的光学读数。缓冲液中 miR-21 的检测具有 0.1 pM 至 1000 pM 的宽线性范围，检测限低至 0.76 pM。幸运的是，该策略证明了稀释血清中 miR-21 检测的性能明显改善。荧光信号明显增强，10%血清中灵敏度进一步提高至0.12 pM。miR-21定量的稳定性和单核苷酸多态性（SNP）分析的能力也大大提高。更重要的是，该生物传感器可用于对不同活体肿瘤细胞中的 miR-21 进行高分辨率成像，说明其在各种复杂情况下检测 miRNA 以进行早期疾病诊断和细胞生物学研究的巨大潜力。
更新日期：2023-10-17
中文翻译：
稳健的非靶标 DNA 触发催化发夹组装扩增策略，用于改善复杂生物基质中 microRNA 的传感
构建了一种基于非靶标 DNA 触发的催化发夹组装 (CHA) 的简单但稳健的荧光策略来探测 microRNA-21 (miR-21)。使用短 ssDNA 作为引发剂，而不是可降解的靶 miRNA。简化了辅助 CHA 过程所需的两个分子信标，以避免不利的非特异性相互作用。在靶标存在的情况下，引发剂从部分双链体中释放出来并触发循环 CHA 反应，从而产生显着放大的光学读数。缓冲液中 miR-21 的检测具有 0.1 pM 至 1000 pM 的宽线性范围，检测限低至 0.76 pM。幸运的是，该策略证明了稀释血清中 miR-21 检测的性能明显改善。荧光信号明显增强，10%血清中灵敏度进一步提高至0.12 pM。miR-21定量的稳定性和单核苷酸多态性（SNP）分析的能力也大大提高。更重要的是，该生物传感器可用于对不同活体肿瘤细胞中的 miR-21 进行高分辨率成像，说明其在各种复杂情况下检测 miRNA 以进行早期疾病诊断和细胞生物学研究的巨大潜力。
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