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An enzyme-free surface plasmon resonance imaging biosensing method for highly sensitive detection of microRNA based on catalytic hairpin assembly and spherical nucleic acid

机译:一种基于催化发夹组装和球形核酸的微小瘤高灵敏度检测的无酶表面等离子体共振成像方法

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摘要

MicroRNAs (miRNAs), considered as therapeutic targets and biomarkers, play important roles in biological processes. Herein, an enzyme-free surface plasmon resonance imaging (SPRi) biosensing method has been developed for miRNA detection based on catalytic hairpin assembly and spherical nucleic acid. The hairpin H1 tethered on the surface of the sensor chip is unfolded by miRNA, and then the hybridized miRNA is released through the displacement of the hairpin H2 for the successive hybridization and assembly process. The emerging DNA fragments on the sensor chip surface after hairpins assembly are further used to hybridize with spherical nucleic acid, inducing a remarkably amplified SPR signal. This biosensing method is highly sensitive to miRNA with a detection limit of 53.7 fM and a linear range of 4 orders of magnitude. Moreover, the biosensor demonstrates good specificity and has the ability to distinguish members of homologous miRNA family even with single base differences. Thus, the SPRi biosensing method may hold a great promise for further application in early clinical diagnosis. (C) 2020 Elsevier B.V. All rights reserved.
机译:MicroRNAS(miRNA)被视为治疗靶标和生物标志物,在生物过程中起重要作用。在此,已经开发了一种基于催化发夹组件和球形核酸的miRNA检测的无酶表面等离子体共振成像(SPRI)生物传感方法。在传感器芯片表面上系的发夹H1由miRNA展开,然后通过发夹H2的位移释放杂交的miRNA以进行连续的杂交和组装过程。在发夹组件之后传感器芯片表面上的新出现的DNA片段进一步用于与球形核酸杂交,诱导显着放大的SPR信号。这种生物抑制方法对miRNA高度敏感,检测限为53.7 fm,线性范围为4个级别。此外,生物传感器展示了良好的特异性,并且能够区分同源miRNA家族的成员,即使是单一的碱差异。因此,SPRI生物体传感方法可以在早期临床诊断中进行进一步应用。 (c)2020 Elsevier B.V.保留所有权利。

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