Homogenous fluorescence polarization assay for the DNA of HIV A T7 by exploiting exonuclease-assisted quadratic recycling amplification and the strong interaction between graphene oxide and ssDNA
Homogenous fluorescence polarization assay for the DNA of HIV A T7 by exploiting exonuclease-assisted quadratic recycling amplification and the strong interaction between graphene oxide and ssDNA
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利用核酸外切酶辅助的二次循环放大以及氧化石墨烯和单链DNA之间的强相互作用对HIV A T7的DNA进行均相荧光偏振测定
DOI：
10.1007/s00604-016-1844-1
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发表时间：
2016-04
影响因子：
5.7
通讯作者：
Zhao Shulin
中科院分区：
文献类型：
--
作者：
Wang Lijun;Tian Jianniao;Huang Yan;Lin Xiaowen;Yang Wen;Zhao Yanchun;Zhao Shulin
作者描述了一种基于荧光偏振 (FP) 的 HIV 检测策略。该测定基于 T7 核酸外切酶辅助二次循环放大，使用氧化石墨烯 (GO) 作为 FP 放大器。这种效应是由于发现 GO 和 ssDNA 之间的结合力比 GO 和 dsDNA 之间的结合力强得多而引起的。由于 5' 末端突出，P1 和染料标记的 P2 两种探针可抵抗 T7 核酸外切酶 (T7exo) 的消化。发夹探针吸附在没有目标 DNA 的 GO 表面。而目标 DNA 的存在会在 T7exo 的帮助下导致两个独立且同时的回收过程。由于这种二次扩增，大量的ssDNA将被吸附到GO表面，这将导致FP的强烈增强。该极化测定用于设计一种测定 HIV DNA 的方法，其检测限为 9.12 pmol·L−1。它具有高度选择性，并且可以在均质溶液中进行。级联扩增策略并不限于这种特定的 DNA，但被认为可以为广谱 DNA 目标分析物的灵敏测定提供通用平台。图解描述了一种荧光偏振策略，该策略基于 T7 核酸外切酶辅助二次循环扩增，并使用氧化石墨烯作为 DNA 的信号放大器工具。
The authors describe a fluorescence polarization (FP) based strategy for HIV detection. The assay is based on T7 exonuclease assisted quadratic recycling amplification using graphene oxide (GO) as an amplifier of FP. The effect is caused by the finding that the binding forces between GO and ssDNA are much stronger than those between GO and dsDNA. The two probes, P1 and dye labeled P2, resist digestion by T7 exonuclease (T7exo) due to the 5′-termini protrusion. The hairpin probes are absorbed on the surface of GO without the target DNA. While the presence of the target DNA leads to two independent and simultaneous recycling processes with the assistance of T7exo. Due to this quadratic amplification, a large quantity of ssDNA will be absorbed onto the surface of GO, and this will cause a strong enhancement of FP. This polarization assay was applied to design a method for the determination of HIV DNA which displays a detection limit of 9.12 pmol·L−1. It is highly selective and can be performed in homogeneous solutions. The cascade amplification strategy is not limited to this particular DNA but is perceived to provide a universal platform for sensitive determination of a wide spectrum of DNA target analytes.Graphical AbstractA fluorescence polarization strategy is described that is based on T7 exonuclease assisted quadratic recycling amplification with graphene oxide acting as a signal amplifier tool for DNA.
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影响因子：
7.4
作者：
Huang, Jiahao;Gao, Xiang;Li, Zhigang
通讯作者：
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影响因子：
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作者：
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通讯作者：
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影响因子：
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作者：
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基于 G-四链体 DNAzyme 的 VEGF165 检测电化学发光生物传感策略：适体-靶标识别和 T7 核酸外切酶辅助循环信号放大的组合
影响因子：
12.6
作者：
Zhang, Hui;Li, Meixing;Cai, Chenxin
通讯作者：
Cai, Chenxin
DOI：
10.1039/c3an00830d
发表时间：
2013-07
期刊：
The Analyst
影响因子：
--
作者：
Juanni Zhang;Jianniao Tian;Yan-Hui He;Sheng Chen;Yixuan Jiang;Yanchun Zhao;Shulin Zhao
通讯作者：
Juanni Zhang;Jianniao Tian;Yan-Hui He;Sheng Chen;Yixuan Jiang;Yanchun Zhao;Shulin Zhao