速度抖动
化学
等位基因
基因分型
打字
摆动碱基对
遗传学
基因型
生物化学
生物
基因
物理
转移RNA
核糖核酸
经典力学
作者
Zhou-Jie Liu,Liang-Yong Yang,Tai-Cheng Lu,Caihong Huang,Yuqi Liang,Xiong-Wei Xu,Yanfang Xu,Mengmeng Liu,Xinhua Lin,Jin-Yuan Chen
标识
DOI:10.1021/acs.analchem.3c01907
摘要
Due to the comparable stability between the perfect-base pair and the wobble-base pair, a precise differentiation of the wobble-type allele has remained a challenge, often leading to false results. Herein, we proposed a ligase chain reaction (LCR)-based ratiometric electrochemical DNA sensor, namely, R-eLCR, for a precise typing of the wobble-type allele, in which the traditionally recognized "negative" signal of wobble-base pair-mediated amplification was fully utilized as a "positive" one and a ratiometric readout mode was employed to ameliorated the underlying potential external influence and improved its detection accuracy in the typing of the wobble-type allele. The results showed that the ratio between current of methylene blue (IMB) and current of ferrocene (IFc) was partitioned in three regions and three types of wobble-type allele were thus precisely differentiated (AA homozygote: IMB/IFc > 2; GG homozygote: IMB/IFc < 1; GA heterozygote: 1 < IMB/IFc < 2); the proposed R-eLCR successfully discriminated the three types of CYP2C19*2 allele in nine cases of human whole blood samples, which was consistent with those of the sequencing method. These results evidence that the proposed R-eLCR can serve as an accurate and robust alternative for the identification of wobble-type allele, which lays a solid foundation and holds great potential for precision medicine.
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