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A dendritically amplified fluorescent signal probe on SiO2 microspheres for the ultrasensitive detection of mercury ions.
Analyst ( IF 3.6 ) Pub Date : 2020/02/10 , DOI: 10.1039/d0an00158a
Chunli Li 1 , Yuqi Zhang , Qianqian Cai , Guifen Jie , Chunxiang Li
Affiliation  

In this work, a new kind of dendritically amplified fluorescent signal probe on SiO2 microspheres was controllably fabricated by the terminal deoxynucleotidyl transferase (TdT)-catalyzed incorporation of nucleotides combined with bio-barcode (BBC) amplification for the ultrasensitive detection of Hg2+. A thymine T-Hg2+-T hairpin structure was first formed and further initiated the strand displacement amplification (SDA) reaction, generating a mimic target (MT). MT hybridized with a capture probe 1 (C1) on SiO2 microspheres, and the 3'-hydroxyl (OH) termini of MT initiated TdT-based DNA extension, producing abundant poly-guanine sequences (G1). Then, G1 hybridized with a capture probe 2 (C2) with abundant cytosine (C) species to assemble multiple C2/reporter probe-AuNPs onto the SiO2 microspheres. The reporter DNA further initiated TdT-based extension with a poly-T sequence (T1) to link large numbers of signal probes, which generated a very high fluorescence signal for the ultrasensitive detection of target Hg2+. This TdT-based signal amplification method coupled with SDA exhibits extraordinary sensitivity for Hg2+ assay with a limit down to 1.0 aM. The proposed highly sensitive fluorescence strategy holds great potential for detecting targets in environmental and biological fields.

中文翻译:

SiO2微球上的树状放大荧光信号探针,用于超灵敏地检测汞离子。

在这项工作中,通过末端脱氧核苷酸转移酶(TdT)催化的核苷酸掺入结合生物条形码(BBC)扩增,对Hg2 +进行超灵敏检测,可控地制备了一种在SiO2微球上的新型树状放大荧光信号探针。首先形成胸腺嘧啶T-Hg2 + -T发夹结构,并进一步引发链置换扩增(SDA)反应,生成模拟靶标(MT)。MT与SiO2微球上的捕获探针1(C1)杂交,MT的3'-羟基(OH)末端启动了基于TdT的DNA延伸,产生了丰富的聚鸟嘌呤序列(G1)。然后,G1与具有丰富胞嘧啶(C)种类的捕获探针2(C2)杂交,以将多个C2 / reporter探针-AuNPs组装到SiO2微球上。报道分子DNA进一步以多T序列(T1)启动基于TdT的延伸,以连接大量信号探针,从而产生了非常高的荧光信号,用于超灵敏地检测目标Hg2 +。这种基于TdT的信号放大方法与SDA结合使用,对Hg2 +分析显示出非凡的灵敏度,其极限低至1.0 aM。提出的高灵敏度荧光策略在检测环境和生物领域的目标方面具有巨大潜力。
更新日期:2020-03-31
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