Artificial Modified Nucleotides for the Electrochemical Detection of Nucleic Acid Amplification Products

IF 1 4区 化学 Q4 CHEMISTRY, ANALYTICAL
E. V. Suprun, S. A. Khmeleva, K. G. Ptitsyn, L. K. Kurbatov, S. P. Radko
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Abstract

The review describes the fundamental electrochemical properties of nucleic acids manifested on solid electrodes, with an emphasis on the spatial structure of macromolecules. The formation of the double helix impedes the contact of the electroactive groups of the nitrogenous bases with the electrode surface, resulting in the disappearance of the analytical signal of deoxyribonucleic acid (DNA). The insufficient electroactivity of the double-stranded DNA is overcome by introducing electrochemically active fragments into the nucleic acid sequence through the polymerase incorporation of chemically modified nucleotides. Currently, an extensive range of artificial nucleotides has been synthesized, which contain various electroactive groups capable of both oxidation and reduction on electrode surfaces at different potentials. Artificial modified nucleotides must exhibit high electrochemical activity while also serving as good substrates for enzymes (polymerases) involved in nucleic acid amplification reactions. Introducing modified nucleotides instead of natural ones into polymerase reactions represents a compromise between the number of labels inserted in one amplicon and the length and quantity of the resulting products. Modified nucleotides find application in the detection of gene mutations and single-nucleotide polymorphisms, nucleic acid sequencing, determination of protein and peptide concentrations, and the detection of pathogenic viruses and bacteria. With the advancement of isothermal amplification methods, the development, synthesis, and investigation of artificial nucleotides have become highly relevant for creating new off-laboratory electrochemical nucleic acid analyzers.

Abstract Image

用于核酸扩增产物电化学检测的人工修饰核苷酸
摘要 本综述介绍了核酸在固体电极上表现出的基本电化学特性,重点是大分子的空间结构。双螺旋的形成阻碍了含氮碱基的电活性基团与电极表面的接触,导致脱氧核糖核酸(DNA)分析信号的消失。双链 DNA 电活性不足的问题可以通过聚合酶在核酸序列中加入化学修饰的核苷酸来克服。目前,已经合成了多种人工核苷酸,它们含有各种电活性基团,在不同电位下能够在电极表面发生氧化和还原反应。人工修饰的核苷酸必须具有较高的电化学活性,同时也是核酸扩增反应中酶(聚合酶)的良好底物。在聚合酶反应中引入修饰核苷酸而不是天然核苷酸,是在一个扩增子中插入的标签数量与所得产物的长度和数量之间的折衷。修饰核苷酸可用于检测基因突变和单核苷酸多态性、核酸测序、测定蛋白质和肽的浓度,以及检测致病病毒和细菌。随着等温扩增方法的发展,人工核苷酸的开发、合成和研究与制造新型实验室外电化学核酸分析仪密切相关。
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来源期刊
Journal of Analytical Chemistry
Journal of Analytical Chemistry 化学-分析化学
CiteScore
2.10
自引率
9.10%
发文量
146
审稿时长
13 months
期刊介绍: The Journal of Analytical Chemistry is an international peer reviewed journal that covers theoretical and applied aspects of analytical chemistry; it informs the reader about new achievements in analytical methods, instruments and reagents. Ample space is devoted to problems arising in the analysis of vital media such as water and air. Consideration is given to the detection and determination of metal ions, anions, and various organic substances. The journal welcomes manuscripts from all countries in the English or Russian language.
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