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一种基于变构球形纳米探针的用于循环微小RNA即时检测的新型定量平台。

A novel quantification platform for point-of-care testing of circulating MicroRNAs based on allosteric spherical nanoprobe.

作者信息

Tian Huiyan, Yuan Changjing, Liu Yu, Li Zhi, Xia Ke, Li Mengya, Xie Fengxin, Chen Qinghai, Chen Ming, Fu Weiling, Zhang Yang

机构信息

Department of Laboratory Medicine, First Affiliated Hospital, Third Military Medical University (Army Medical University), Chongqing, China.

Department of Laboratory Medicine, Chongqing University Cancer Hospital, Chongqing, China.

出版信息

J Nanobiotechnology. 2020 Oct 31;18(1):158. doi: 10.1186/s12951-020-00717-z.

Abstract

MiRNA-150, a gene regulator that has been revealed to be abnormal expression in non-small cell lung cancer (NSCLC), can be regarded as a serum indicator for diagnosis and monitoring of NSCLC. Herein, a new sort of nanoprobe, termed allosteric spherical nanoprobe, was first developed to sense miRNA-150. Compared with conventional hairpin, this new nanoprobe possesses more enrichment capacity and reaction cross section. Structurally, it consists of magnetic nanoparticles and dual-hairpin. In the absence of miRNA-150, the spherical nanoprobes form hairpin structure through DNA self-assembly, which could promote the Förster resonance energy transfer (FRET) of fluorophore (FAM) and quencher (BHQ1) nearby. However, in the presence of target, the target-probe hybridization can open the hairpin and form the active "Y" structure which separated fluorophore and quencher to yield "signal on" fluorescence. In the manner of multipoint fluorescence detection, the target-bound allosteric spherical nanoprobe could provide high detection sensitivity with a linear range of 100 fM to 10 nM and a detection limit of 38 fM. More importantly, the proposed method can distinguish the expression of serum miRNA-150 among NSCLC patients and healthy people. Finally, we hoped that the potential bioanalytical application of this nanoprobe strategy will pave the way for point-of-care testing (POCT).

摘要

微小RNA-150是一种在非小细胞肺癌(NSCLC)中已被揭示存在异常表达的基因调节因子,可被视为用于NSCLC诊断和监测的血清指标。在此,首次开发了一种新型纳米探针,称为变构球形纳米探针,用于检测微小RNA-150。与传统发夹相比,这种新型纳米探针具有更强的富集能力和反应截面。在结构上,它由磁性纳米颗粒和双发夹组成。在不存在微小RNA-150的情况下,球形纳米探针通过DNA自组装形成发夹结构,这可以促进附近荧光团(FAM)和猝灭剂(BHQ1)的荧光共振能量转移(FRET)。然而,在有靶标的情况下,靶标与探针的杂交可以打开发夹并形成活性“Y”结构,从而使荧光团和猝灭剂分离,产生“信号开启”荧光。以多点荧光检测的方式,与靶标结合的变构球形纳米探针可以提供高检测灵敏度,线性范围为100 fM至10 nM,检测限为38 fM。更重要的是,所提出的方法可以区分NSCLC患者和健康人血清中微小RNA-150的表达。最后,我们希望这种纳米探针策略的潜在生物分析应用将为即时检测(POCT)铺平道路。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5a5b/7603675/93d11ef80f8a/12951_2020_717_Sch1_HTML.jpg

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