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基于双链特异性核酸酶触发滚环扩增构建微小RNA-21太赫兹超材料传感方法

[Construction of microRNA-21 terahertz metamaterial sensing method based on duplex-specific nuclease triggered rolling circle amplification].

作者信息

Zhan X Y, Yang S, Zhang Y, Yang X, Fu W L

机构信息

Department of Clinical Laboratory Medicine, Southwest Hospital, Army Medical University, Chongqing 400038, China.

出版信息

Zhonghua Yu Fang Yi Xue Za Zhi. 2021 Feb 6;55(2):212-218. doi: 10.3760/cma.j.cn112150-20201111-01359.

Abstract

To construct of a terahertz metamaterial sensing method for detection of microRNA-21 (miRNA-21) based on duplex-specific nuclease triggered rolling circle amplification. First, a THz metamaterial sensing method was constructed, and verified by polyacrylamide gel electrophoresis and zeta potential; after optimizing the detection conditions of the sensor, different concentrations of miRNA-21 and other different miRNAs were detected. And this biosensor was compared with other miRNA detection methods; finally, the recovery rate of the biosensor was evaluated. Under the optimal experimental conditions, through the dual signal amplification strategy of duplex-specific nuclease (DSN) cycle recognition and rolling circle amplification (RCA), the THz metamaterial sensor has a response range of 10 fmol/L to 10 nmol/L to the target miRNA-21, with a detection limit of 8.49 fmol/L. And the biosensor has good specificity with the ability to recognize the target miRNA-21 from a variety of microRNAs. And the experiment with recovery rate from 94.33% to 115.33% has been further verified in commercial human serum samples. The terahertz biosensor can achieve highly sensitive and specific detection of the target miRNA-21, which proves the potential for label-free diagnosis and early warning of miRNA-related diseases.

摘要

构建一种基于双链特异性核酸酶触发滚环扩增的太赫兹超材料传感方法用于检测微小RNA-21(miRNA-21)。首先,构建了一种太赫兹超材料传感方法,并通过聚丙烯酰胺凝胶电泳和zeta电位进行验证;在优化传感器检测条件后,检测了不同浓度的miRNA-21和其他不同的微小RNA。并将该生物传感器与其他miRNA检测方法进行比较;最后,评估了该生物传感器的回收率。在最佳实验条件下,通过双链特异性核酸酶(DSN)循环识别和滚环扩增(RCA)的双信号放大策略,太赫兹超材料传感器对目标miRNA-21的响应范围为10 fmol/L至10 nmol/L,检测限为8.49 fmol/L。并且该生物传感器具有良好的特异性,能够从多种微小RNA中识别目标miRNA-21。在商业人血清样本中进一步验证了回收率在94.33%至115.33%的实验。太赫兹生物传感器能够实现对目标miRNA-21的高灵敏和特异性检测,这证明了其在miRNA相关疾病的无标记诊断和早期预警方面的潜力。

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