A two-layer assay for single-nucleotide variants utilizing strand displacement and selective digestion | |
Yu, Yingjie ; Wu, Tongbo ; Johnson-Buck, Alexander ; Li, Lidan ; Su, Xin | |
刊名 | BIOSENSORS & BIOELECTRONICS |
2016 | |
关键词 | Single nucleotide variant Toehold strand displacement Lambda exonuclease Fluorescent probe DNA HYBRIDIZATION MUTATIONS MEDICINE PLASMA |
DOI | 10.1016/j.bios.2016.03.070 |
英文摘要 | Point mutations have emerged as prominent biomarkers for disease diagnosis, particularly in the case of cancer. Discovering single-nucleotide variants (SNVs) is also of great importance for the identification of single-nucleotide polymorphisms within the population. The competing requirements of thermodynamic stability and specificity in conventional nucleic acid hybridization probes make it challenging to achieve highly precise detection of point mutants. Here, we present a fluorescence-based assay for low abundance mutation detection based on toehold-mediated strand displacement and nuclease-mediated strand digestion that enables highly precise detection of point mutations. We demonstrate that this combined assay provides 50-1000-fold discrimination (mean value: 255) between all possible single nucleotide mutations and their corresponding wild-type sequence for a model DNA target. Using experiments and kinetic modeling, we investigate probe properties that obtain additive benefits from both strand displacement and nucleolytic digestion, thus providing guidance for the design of enzyme mediated nucleic acid assays in the future. (C) 2016 Elsevier B.V. All rights reserved.; National Natural Science Foundation of China [81273631]; subject promotion plan of Beijing University of Chemical Technology [ys12060026025]; SCI(E); EI; PubMed; ARTICLE; xinsu@mail.buct.edu.cn; 248-254; 82 |
语种 | 英语 |
内容类型 | 期刊论文 |
源URL | [http://ir.pku.edu.cn/handle/20.500.11897/434028] |
专题 | 化学与分子工程学院 |
推荐引用方式 GB/T 7714 | Yu, Yingjie,Wu, Tongbo,Johnson-Buck, Alexander,et al. A two-layer assay for single-nucleotide variants utilizing strand displacement and selective digestion[J]. BIOSENSORS & BIOELECTRONICS,2016. |
APA | Yu, Yingjie,Wu, Tongbo,Johnson-Buck, Alexander,Li, Lidan,&Su, Xin.(2016).A two-layer assay for single-nucleotide variants utilizing strand displacement and selective digestion.BIOSENSORS & BIOELECTRONICS. |
MLA | Yu, Yingjie,et al."A two-layer assay for single-nucleotide variants utilizing strand displacement and selective digestion".BIOSENSORS & BIOELECTRONICS (2016). |
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