Detection of single nucleotide polymorphism by measuring extension kinetics with T7 exonuclease mediated isothermal amplification,Analyst

当前位置： X-MOL 学术 › Analyst › 论文详情

Our official English website, www.x-mol.net, welcomes your feedback! (Note: you will need to create a separate account there.)

Detection of single nucleotide polymorphism by measuring extension kinetics with T7 exonuclease mediated isothermal amplification
Analyst ( IF 3.6 ) Pub Date : 2017-10-17 00:00:00 , DOI: 10.1039/c7an00875a
Miao Cui _{1,

2,

3,

4,

5} , Xianjin Xiao _{5,

6,

7,

8,

9} , Meiping Zhao _{10,

11,

12,

13,

14} , Bo Zheng _{1,

2,

3,

4,

5}

Affiliation

In this work, we measured the primer extension kinetics of the Klenow fragment (exo-) to achieve rapid detection of single nucleotide polymorphism (SNP). Both the matching and the single-base mismatching targets were used as the primer in the kinetic measurements to identify the single nucleotide polymorphism. By coupling with the T7 exonuclease-assisted target cycling process, we decreased the detection limit but still maintained a high discrimination factor. After the demonstration of a good discrimination ability with synthetic DNA strands, we applied the method to detect low abundance of epidermal growth factor receptor (EGFR) mutation in human genomic DNA, which was a biomarker of non-small cell lung cancer. The kinetics based SNP detection was performed at room temperature and was robust against photobleaching and other optical interferences for the detection of low abundance of point mutations in human genomic DNA. The detection method is adaptable to a microarray platform for high-throughput and point-of-care detection.

中文翻译：

通过测量T7核酸外切酶介导的等温扩增的延伸动力学来检测单核苷酸多态性

在这项工作中，我们测量了Klenow片段的引物延伸动力学（exo-）实现单核苷酸多态性（SNP）的快速检测。匹配和单碱基错配靶都被用作动力学测量中的引物，以鉴定单核苷酸多态性。通过与T7核酸外切酶辅助的靶标循环过程相结合，我们降低了检测限，但仍保持了较高的判别率。在证明合成DNA链具有良好的辨别能力后，我们应用该方法检测了人类基因组DNA中表皮生长因子受体（EGFR）突变的低丰度，这是非小细胞肺癌的生物标记。基于动力学的SNP检测是在室温下进行的，对于检测人类基因组DNA中的点突变的低丰度，具有很强的抗光漂白和其他光学干扰的能力。

更新日期：2017-11-08

点击分享查看原文

点击收藏

阅读更多本刊最新论文本刊介绍/投稿指南11