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基于催化发夹组装和金纳米棒刻蚀的端粒酶活性灵敏多色可视化检测。

Sensitive multicolor visual detection of telomerase activity based on catalytic hairpin assembly and etching of Au nanorods.

机构信息

Research Center for Analytical Sciences, Northeastern University, Shenyang 110819, PR China.

Research Center for Analytical Sciences, Northeastern University, Shenyang 110819, PR China.

出版信息

Biosens Bioelectron. 2018 Dec 30;122:247-253. doi: 10.1016/j.bios.2018.09.064. Epub 2018 Sep 19.

Abstract

Detection of telomerase activity is crucial for the telomerase-related early diagnosis of cancer and drug screening. Herein, a multicolor visual telomerase detection method with high sensitivity was developed based on the etching of hexadecyl trimethyl ammonium bromide-stablized Au nanorods (Au NRs) by the oxidation state 3,3',5,5'-tetramethylbenzidine sulfate (TMB). In order to meet the demand of bare-eye inspection, an enzyme-free signal amplification strategy of catalytic hairpin assembly (CHA) was incorporated. After the introduction of telomerase, telomerase extension products specifically triggered cyclic CHA and led to the successive formation of G-quadruplex/hemin DNAzyme, which catalyzed the HO-mediated oxidation of TMB. The Au NRs were gradually etched as the concentration of catalysate TMB increased, resulting in the decrease of aspect ratio of the Au NRs. Correspondingly, the longitudinal localized surface plasmon resonance peak of Au NRs was blue-shifted, with the concomitant generation of a series of color transition. Under optimal conditions, a highly sensitive detection toward telomerase was realized down to 15 HeLa cells. Compared to previous colorimetric method for telomerase determination, multiple colors corresponding to telomerase activity was the most attractive virtue of our approach. More strikingly, telomerase-induced cyclic CHA significantly enhanced the accumulation of G-quadruplex/hemin DNAzyme, thus the signal amplification was effectively realized. Our approach exhibited excellent sensitivity and convenient signal readout, which is expected to provide great potential application in cancer diagnosis and therapy.

摘要

端粒酶活性的检测对于基于端粒酶的癌症早期诊断和药物筛选至关重要。在此,我们基于 3,3',5,5'-四甲基联苯胺硫酸盐(TMB)的氧化态对十六烷基三甲基溴化铵稳定的 Au 纳米棒(Au NRs)进行刻蚀,开发了一种高灵敏度的多色可视化端粒酶检测方法。为了满足肉眼观察的需求,我们引入了无酶信号放大策略——催化发夹组装(CHA)。在引入端粒酶后,端粒酶延伸产物特异性触发循环 CHA,并导致 G-四链体/血红素 DNA 酶的连续形成,从而催化 TMB 的 HO 介导氧化。随着催化产物 TMB 浓度的增加,Au NRs 逐渐被刻蚀,导致 Au NRs 的纵横比降低。相应地,Au NRs 的纵向局域表面等离激元共振峰发生蓝移,同时产生一系列颜色转变。在最佳条件下,我们实现了对端粒酶的高灵敏度检测,检测下限低至 15 个 HeLa 细胞。与之前用于端粒酶测定的比色法相比,我们的方法最吸引人的优点是对应于端粒酶活性的多种颜色。更引人注目的是,端粒酶诱导的循环 CHA 显著增强了 G-四链体/血红素 DNA 酶的积累,从而有效地实现了信号放大。我们的方法表现出优异的灵敏度和方便的信号读出,有望在癌症诊断和治疗中提供巨大的潜在应用价值。

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