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Nanographite-based fluorescent biosensor for detecting microRNA using duplex-specific nuclease-assisted recycling.
Luminescence ( IF 3.2 ) Pub Date : 2019-12-16 , DOI: 10.1002/bio.3733
Qizhi He 1 , Huaiqing Luo 1 , Lingli Chen 2 , Jun Dong 1 , Keke Chen 1 , Yi Ning 2
Affiliation  

The development of a nanographite (NG)-based fluorescent biosensor for detecting microRNA (miRNA) is reported. Duplex-specific nuclease (DSN)-assisted signal amplification was key to its function. In the absence of a target, with the assistance of p-stacking interactions, the NG adsorbed the double carboxyfluorescein (FAM)-labelled probe (DFP) whose surface was perfectly complementary to miRNA, leading to quenching of FAM fluorescence. In the presence of a target, double-stranded DNA/RNA hybrids were repelled by the NG and fluorescence was restored. Meanwhile, the considerable increase in signal strength and sensitivity suggests DSN-mediated target recycling as an application. The detection limit of the proposed biosensor for miRNA was 10 pmol/L; there was a linear correlation when the miRNA concentration ranged from 50 pmol/L to 5 nmol/L. Additionally, the method could distinguish let-7b from most let-7 miRNA family members and was successfully used in a sample assay. This biosensor is a novel and highly sensitive tool for miRNA detection and has great potential for biochemical research, disease diagnosis, and therapy.

中文翻译:

基于纳米石墨的荧光生物传感器,用于使用双特异性特异性核酸酶辅助回收检测microRNA。

据报道,用于检测微小RNA(miRNA)的基于纳米石墨(NG)的荧光生物传感器的发展。双链特异性核酸酶(DSN)辅助信号放大是其功能的关键。在没有靶标的情况下,借助p堆积相互作用,NG吸附了表面与miRNA完全互补的双羧基荧光素(FAM)标记的探针(DFP),导致FAM荧光淬灭。在存在靶标的情况下,NG排斥双链DNA / RNA杂种,并恢复了荧光。同时,信号强度和灵敏度的显着提高表明DSN介导的靶标回收是一种应用。拟议的miRNA生物传感器的检出限为10 pmol / L;当miRNA浓度在50 pmol / L至5 nmol / L之间时,存在线性相关。另外,该方法可以将let-7b与大多数let-7 miRNA家族成员区分开,并成功用于样品测定中。这种生物传感器是用于miRNA检测的新型且高度灵敏的工具,在生化研究,疾病诊断和治疗方面具有巨大潜力。
更新日期:2020-04-03
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