Highly sensitive and selective chemiluminescent imaging for DNA detection by ligation-mediated rolling circle amplified synthesis of DNAzyme
Highly sensitive and selective chemiluminescent imaging for DNA detection by ligation-mediated rolling circle amplified synthesis of DNAzyme
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DOI：
10.1016/j.bios.2012.08.050
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发表时间：
2013-03-15
影响因子：
12.6
通讯作者：
Zhang, Xueji
中科院分区：
文献类型：
--
作者：
Dong, Haifeng;Wang, Chen;Zhang, Xueji
通过设计一种模拟DNAzyme的辣根过氧化物酶的放大合成方法，并将其引入化学发光(CL)成像，提出了一种高灵敏度、高选择性的亚毫微摩尔水平的DNA生物传感方法。扩增合成是通过结合靶DNA连接酶反应和滚环扩增(RCA)实现的，滚环扩增产生数千个重复序列与氯化血红素结合，形成大量辣根过氧化物酶DNAzyme单位。该放大策略大大提高了鲁米诺-双氧水体系的化学发光强度。该基因分型方法在等位基因识别方面表现出高度的特异性。基于连接介导的DNAzyme RCA合成的化学发光成像新策略在区分单碱基错配方面表现出高度的保真度，并有效地促进了用于敏感靶DNA检测的信号放大。检测范围为1×10~(-15)M~1×10~(-11)M，检出限为0.26 fM。所提出的方法为生物分析和临床生物医学应用中的遗传靶点分析提供了一个健壮、经济、高度敏感和特异的平台。(C)2012爱思唯尔B.V.保留所有权利。
A highly sensitive DNA biosensing method down to sub-femtomolar level with excellent selectivity was proposed by designing an amplified synthesis of horseradish peroxidase mimicking DNAzyme and introducing the amplified DNAzyme to chemiluminescent (CL) imaging. The amplified synthesis was achieved by combining a target DNA related ligase reaction with rolling circle amplification (RCA), which produced thousands of repeated sequences to bind hemin and form a mass of horseradish peroxidase-mimicing DNAzyme units. The amplification strategy greatly enhanced the CL emission of the luminol-H2O2 system. The genotyping method displayed highly specific biochemistry in allele discrimination. The novel CL imaging strategy based on ligation-mediated RCA synthesis of DNAzyme showed high fidelity in discriminating single-base mismatch and efficiently facilitated signal amplification for sensitive target DNA detection. It could detect DNA ranging from 1 x 10(-15) M to 1 x 10(-11) M with a detection limit of 0.26 fM. The proposed approach provided a robust, cost-efficient, highly sensitive and specific platform for genetic target analysis in bioanalysis and clinic biomedical application. (C) 2012 Elsevier B.V. All rights reserved.