title: Label-free genotyping of cytochrome P450 2D6*10 using ligation-mediated strand displacement amplification with DNAzyme-based chemiluminescence detection.
abstract: 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.
authors: State Key Laboratory of Chemo/Bio-Sensing and Chemometrics, College of Chemistry and Chemical Engineering, Hunan University, Changsha, PR China.
authors: Hong-Qi
authors: HQ
authors: Wang
authors: Zhan
authors: Z
authors: Wu
authors: Yan
authors: Y
authors: Zhang
authors: Li-Juan
authors: LJ
authors: Tang
authors: Ru-Qin
authors: RQ
authors: Yu
authors: Jian-Hui
authors: JH
authors: Jiang
mesh_terms: D052157:Aptamers, Nucleotide / Q000737:chemistry; D020137:Base Pair Mismatch; D019389:Cytochrome P-450 CYP2D6 / Q000235:genetics*; D000072481:DNA Ligase ATP; D011088:DNA Ligases / Q000378:metabolism; D021881:DNA, Catalytic / Q000378:metabolism*; D005838:Genotype; D006427:Hemin / Q000737:chemistry; D008163:Luminescent Measurements; D021141:Nucleic Acid Amplification Techniques; D020641:Polymorphism, Single Nucleotide
chemical_list: D052157:Aptamers, Nucleotide; D021881:DNA, Catalytic; D006427:Hemin; D019389:Cytochrome P-450 CYP2D6; D011088:DNA Ligases; D000072481:DNA Ligase ATP