Label-free genotyping of cytochrome P450 2D6*10 using ligation-mediated strand displacement amplification with DNAzyme-based chemiluminescence detection

文献类型： 外文期刊

第一作者： Wang, Hong-Qi

作者： Wang, Hong-Qi;Wu, Zhan;Zhang, Yan;Tang, Li-Juan;Yu, Ru-Qin;Jiang, Jian-Hui;Wang, Hong-Qi

作者机构：

关键词： Genotyping;Cytochrome P450 monooxygenase 2D6*10;DNA ligase;Strand displacement amplification;Single nucleotide polymorphism

期刊名称：ANALYTICA CHIMICA ACTA （ 影响因子：6.558； 五年影响因子：6.228 ）

ISSN： 0003-2670

年卷期： 2012 年 710 卷

页码：

收录情况： SCI

摘要： Genotyping of cytochrome P450 monooxygenase 2D6*10 (CYP2D6*10) plays an important role in pharmacogenomics, especially in clinical drug therapy of Asian populations. This work reported a novel label-free technique for genotyping of CYP2D6*10 based on ligation-mediated strand displacement amplification (SDA) with DNAzyme-based chemiluminescence detection. Discrimination of single-base mismatch is firstly accomplished using DNA ligase to generate a ligation product. The ligated product then initiates a SDA reaction to produce aptamer sequences against hemin, which can be probed by chemiluminescence detection. The proposed strategy is used for the assay of CYP2D6*10 target and the genomic DNA. The results reveal that the proposed technique displays chemiluminescence responses in linear correlation to the concentrations of DNA target within the range from 1 pM to 1 nM. A detection limit of 0.1 pM and a signal-to-background ratio of 57 are achieved. Besides such high sensitivity, the proposed CYP2D6*10 genotyping strategy also offers superb selectivity, great robustness, low cost and simplified operations due to its label-free, homogeneous, and chemiluminescence-based detection format. These advantages suggest this technique may hold considerable potential for clinical CYP2D6*10 genotyping and association studies. (C) 2011 Elsevier B.V. All rights reserved.

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