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利用硅纳米线生物传感器对微小RNA进行无标记直接检测。

Label-free direct detection of MiRNAs with silicon nanowire biosensors.

作者信息

Zhang Guo-Jun, Chua Jay Huiyi, Chee Ru-Ern, Agarwal Ajay, Wong She Mein

机构信息

Institute of Microelectronics, Agency for Science, Technology and Research, 11 Science Park Road, 117685 Singapore, Singapore.

出版信息

Biosens Bioelectron. 2009 Apr 15;24(8):2504-8. doi: 10.1016/j.bios.2008.12.035. Epub 2009 Jan 4.

Abstract

MicroRNA (miRNA), an 18-24-nucleotide (nt) noncoding RNA molecule in the genes of humans, plants and animals, is emerging as a key player in gene regulation. As a result, label-free, rapid, and sensitive detection for miRNA is of great significance. In this work, a label-free and direct hybridization assay for ultrasensitive detection of miRNA using silicon nanowires (SiNWs) device has been developed. Peptide nucleic acids (PNAs), which serve as a receptor to recognize miRNA directly without labeling the target miRNA, are immobilized on the surface of the SiNW device. Resistance change measured before and after hybridization correlates directly to concentrations of the hybridized target miRNA. Concentration-dependent measurements indicate that a detection limit of 1 fM was obtained using the optimized assay. The technique enables identification of fully matched versus mismatched miRNA sequences. Furthermore, the SiNW device is capable of detecting miRNA in total RNA extracted from Hela cells. This approach paves a way for label-free, early detection of miRNA as a biomarker in cancer diagnostics with very high sensitivity and good specificity.

摘要

微小RNA(miRNA)是人类、植物和动物基因中的一种18 - 24个核苷酸(nt)的非编码RNA分子,正逐渐成为基因调控中的关键角色。因此,对miRNA进行无标记、快速且灵敏的检测具有重要意义。在这项工作中,已开发出一种使用硅纳米线(SiNWs)器件对miRNA进行超灵敏检测的无标记直接杂交检测方法。肽核酸(PNA)作为直接识别miRNA的受体,无需对目标miRNA进行标记,被固定在SiNW器件表面。杂交前后测量的电阻变化与杂交目标miRNA的浓度直接相关。浓度依赖性测量表明,使用优化后的检测方法可获得1 fM的检测限。该技术能够区分完全匹配与错配的miRNA序列。此外,SiNW器件能够检测从Hela细胞中提取的总RNA中的miRNA。这种方法为在癌症诊断中以非常高的灵敏度和良好的特异性无标记早期检测miRNA作为生物标志物铺平了道路。

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