Target-triggered autocatalytic sequence recycling for sensitive and simultaneous detection of microRNA and mRNA via multi-donor iFRET signal amplification
Target-triggered autocatalytic sequence recycling for sensitive and simultaneous detection of microRNA and mRNA via multi-donor iFRET signal amplification
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DOI：
10.1016/j.snb.2021.130351
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发表时间：
2021-10
影响因子：
8.4
通讯作者：
Xiaolong Li;Wenjiao Zhou;Lei Liao;Yun Xiang;Bingying Jiang;R. Yuan
中科院分区：
文献类型：
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作者：
Xiaolong Li;Wenjiao Zhou;Lei Liao;Yun Xiang;Bingying Jiang;R. Yuan
同时监测不同的生物标志物分子可以为疾病诊断提供高精度。在这项工作中，通过使用新的生物自动循环邻近记录（APR）方法和多供体诱导荧光共振能量转移（iFRET），我们建立了一种多重且灵敏的荧光方法，用于同时检测microRNA-155和骨桥蛋白mRNA，以准确诊断酒精性肝病。两个靶序列的存在通过两个立足点介导的链置换反应触发 APR 过程，从而生成许多 dsDNA，其中 ROX 和 Cy5 染料连接到其末端。随后，插入 dsDNA 链的 SYBR Green I 染料的激发会通过 iFRET 在不同波长下表现出放大的荧光，从而灵敏且同时检测 microRNA-155 和骨桥蛋白 mRNA。通过 APR 和 iFRET 的协同信号放大，我们的方法显示了两种 RNA 序列的亚飞摩尔检测限（例如，microRNA-155 和骨桥蛋白 mRNA 分别为 0.5 和 0.3 fM）。此外，该方法还可以实现高选择性并检测稀释血清样本中的两条RNA序列，这表明其在其他疾病的准确诊断方面具有潜在的应用前景。
Simultaneous monitoring of different biomarker molecules can potentially offer high accuracy for disease diagnosis. In this work, by using a new biological auto-cycling proximity recording (APR) approach and multi-donor-induced fluorescence resonance energy transfer (iFRET), we establish a multiplexed and sensitive fluorescent method for simultaneous detection of microRNA-155 and osteopontin mRNA for accurate alcoholic liver disease diagnosis. The presence of the two target sequences triggers the APR processviatwo toehold-mediated strand displacement reactions for the generation of many dsDNAs with the ROX and Cy5 dyes linked to their termini. Subsequent excitation of the SYBR Green I dye intercalated into the dsDNA strands exhibits amplified fluorescence at distinct wavelengthsviaiFRET for sensitive and simultaneous detection of microRNA-155 and osteopontin mRNA. With the synergistic signal amplification by APR and iFRET, our method shows sub-femtomolar detection limits for the two RNA sequences (e.g., 0.5 and 0.3 fM for microRNA-155 and osteopontin mRNA, respectively). In addition, such a method can also realize high selectivity and the detection of the two RNA sequences in diluted serum samples, indicating its potential application for accurate diagnosis of other diseases.