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基于核酸内切酶IV的竞争性DNA探针分析法,通过区分稳定的单碱基错配来鉴别低丰度点突变。

Endonuclease IV based competitive DNA probe assay for differentiation of low-abundance point mutations by discriminating stable single-base mismatches.

作者信息

Xu Jiaju, Li Longjie, Chen Na, She Yongxin, Wang Shanshan, Liu Na, Xiao Xianjin

机构信息

Family Planning Research Institute/Center of Reproductive Medicine, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, P. R. China.

出版信息

Chem Commun (Camb). 2017 Aug 22;53(68):9422-9425. doi: 10.1039/c7cc04816e.

Abstract

We disclosed the unique discrimination property of Endo IV toward stable single-base mismatches located at the second nucleotide 3' to the AP site. Coupled with thermodynamic differentiation and competitive blocker strands, a highly sensitive and specific detection system was established with discrimination factors of 510-1079 for G:X mismatches and LODs of 0.003-0.005% for KRAS G12A, KRAS G12V and KRAS G12S mutations.

摘要

我们揭示了内切酶IV对位于脱嘌呤嘧啶位点(AP位点)3'端第二个核苷酸处的稳定单碱基错配的独特识别特性。结合热力学差异和竞争性阻断链,建立了一种高度灵敏且特异的检测系统,对G:X错配的识别因子为510 - 1079,对KRAS G12A、KRAS G12V和KRAS G12S突变的检测限为0.003 - 0.005%。

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