Colorimetric detection of cholesterol with G-quadruplex-based DNAzymes and ABTS2-
Colorimetric detection of cholesterol with G-quadruplex-based DNAzymes and ABTS2-
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DOI：
10.1016/j.aca.2012.02.015
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发表时间：
2012-04-29
影响因子：
6.2
通讯作者：
Ling, Liansheng
中科院分区：
文献类型：
--
作者：
Li, Ruimin;Xiong, Cen;Ling, Liansheng
用血红素- g -四联DNAzyme将胆固醇的氧化转化为2,2'-氮基-双（3-乙基苯并噻唑-6-磺酸）二铵盐（ABTS(2-)）的颜色变化，建立了一种新的检测胆固醇的比色法。寡核苷酸5′- gtgggtagggcgggttgg -3′（Oligo-1）在K+存在下形成g -四重结构，在结合血红素时充当模拟DNAzyme的辣根过氧化物酶（HRP），并催化胆固醇氧化酶催化胆固醇与氧反应产生的H2O2将无色ABTS（2-）氧化为绿色ABTS（中心点-）。因此，将这两种反应结合起来，可以将胆固醇的氧化转化为ABTS（2-）的颜色变化。在最佳条件下，吸光度与胆固醇浓度在1.0 ~ 30 μ M范围内成正比，线性回归方程为a = 0.362 + 0.0256C (C: μ M, R = 0.998)，检出限为0.101 μ M （3 σ /斜率）。此外，还研究了该方法在人血清胆固醇检测中的实用性。(C) 2012 Elsevier B.V.版权所有
A novel colorimetric method for detection of cholesterol was developed with hemin-G-quadruplex DNAzyme by transducing oxidation of cholesterol into the color change of 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt (ABTS(2-)). Oligonucleotide 5'-GTGGGTAGGGCGGGTTGG-3' (Oligo-1) formed G-quadruplex structure in the presence of K+, it acted as a horseradish peroxidase (HRP) mimicking DNAzyme when binding hemin and catalyzed the oxidation of colorless ABTS(2-) to green ABTS(center dot-) by H2O2, which was produced by the reaction of cholesterol and oxygen that catalyzed by cholesterol oxidase. Therefore, the oxidation of cholesterol could be transduced into the color change of ABTS(2-) by combining these two reactions. Under the optimum conditions, the absorbance was proportional to the concentration of cholesterol over the range of 1.0-30 mu M, with a linear regression equation of A= 0.362 + 0.0256C (C: mu M, R = 0.998) and a detection limit of 0.101 mu M (3 sigma/slope). Moreover, the practicability of the assay in the detection of cholesterol in human serum was studied as well. (C) 2012 Elsevier B.V. All rights reserved.