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
作者: Wang, Hong-Qi 1 ; Wu, Zhan 1 ; Zhang, Yan 1 ; Tang, Li-Juan 1 ; Yu, Ru-Qin 1 ; Jiang, Jian-Hui 1 ;
作者机构: 1.Hunan Univ, Coll Chem & Chem Engn, State Key Lab Chemo Biosensing & Chemometr, Changsha 410082, Hunan, Peoples R China
2.Henan Acad Agr Sci, Res Ctr Agr Qual Stand & Testing Tech, Zhengzhou 450002, Peoples R China
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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