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DNA polymerase/NEase-assisted signal amplification coupled with silver nanoclusters for simultaneous detection of multiple microRNAs and molecular logic operations
Sensors and Actuators B: Chemical ( IF 8.0 ) Pub Date : 2020-09-17 , DOI: 10.1016/j.snb.2020.128915
Mei Li , Xiong Xu , Qingyou Cai

Herein, using a novel bifunctional DNA-hybridization-DNA (DHD) probe, a symmetric DNA polymerase/NEase-assisted target signal amplification reaction into a single probe coupled with dual “light-up” colored silver nanoclusters (AgNCs) was introduced for label-free simultaneous detection of multiple microRNAs (miRNAs) for the first time. The DHD probe contained two single-stranded loops complementary to two different miRNA targets. Upon the recognition of the specific target miRNAs (miR-21 or miR-141), a symmetric polymerase/NEase-assisted signal amplification reaction was initiated, which generated massive triggers (trigger 1 and trigger 2). Subsequently, the released triggers hybridized with two different dark AgNCs and formed a partial double-stranded structure; thus, “light-up” red or yellow fluorescence readout was observed. The established biosensing approach was capable of simultaneously detecting multiple miRNAs with a limit of detection (LOD) down to 9.8/6.1 pM for miR-21/miR-141, respectively. This innovative approach displayed acceptable reproducibility and high specificity. Furthermore, the “AND” logic gate was activated by combining the dual-functional DHD probe with the dual “light-up” colored AgNCs, with/without miRNAs as inputs.



中文翻译:

DNA聚合酶/ NEase辅助信号放大与银纳米团簇结合,可同时检测多种microRNA和分子逻辑操作

在本文中,使用新型的双功能DNA杂交DNA(DHD)探针,将对称DNA聚合酶/ NEase辅助的目标信号扩增反应引入与双“亮”色的银纳米簇(AgNC)耦合的单个探针中首次免费同时检测多个microRNA(miRNA)。DHD探针包含两个与两个不同miRNA靶标互补的单链环。识别出特定的靶标miRNA(miR-21或miR-141)后,就启动了对称聚合酶/ NEase辅助信号放大反应,产生了大量的触发信号(触发信号1和触发信号2)。随后,释放的触发器与两种不同的深色AgNC杂交并形成部分双链结构。因此,观察到“点亮”的红色或黄色荧光读数。建立的生物传感方法能够同时检测多种miRNA,而miR-21 / miR-141的检测限(LOD)分别为9.8 / 6.1 pM。这种创新方法显示出可接受的重现性和高特异性。此外,通过将双功能DHD探针与双色“点亮”彩色AgNC结合使用,可以将“ AND”逻辑门激活,无论是否输入miRNA。

更新日期:2020-10-11
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