Specificity Enhancement of Deoxyribonucleic Acid Polymerization for Sensitive Nucleic Acid Detection,Analytical Chemistry

当前位置： X-MOL 学术 › Anal. Chem. › 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.)

Specificity Enhancement of Deoxyribonucleic Acid Polymerization for Sensitive Nucleic Acid Detection
Analytical Chemistry ( IF 6.7 ) Pub Date : 2020-11-25 , DOI: 10.1021/acs.analchem.0c03223
Bei Hu ₁ , Yitao Wang ₁ , Shichao Sun ₁ , Guangcheng Luo ₁ , Shun Zhang ₁ , Jun Zhang ₁ , Lu Chen ₂ , Zhen Huang _{1,

2}

Affiliation

Specificity of DNA polymerization plays a critical role in DNA replication and storage of genetic information. Likewise, biotechnological applications, such as nucleic acid detection, DNA amplification, and gene cloning, require high specificity in DNA synthesis catalyzed by DNA polymerases. However, errors in DNA polymerization (such as mis-incorporation and mis-priming) can significantly jeopardize the specificity. Herein, we report our discovery that the specificity of DNA enzymatic synthesis can be substantially enhanced (up to 100-fold higher) by attenuating DNA polymerase kinetics via the phosphorothioate dNTPs. This specificity enhancement allows convenient and sensitive nucleic acid detection, polymerization, PCR, and gene cloning with complex systems (such as human cDNA and genomic DNA). Further, we found that the specificity enhancement offered higher sensitivity (up to 50-fold better) for detecting nucleic acids, such as COVID-19 viral RNAs. Our findings have revealed a simple and convenient strategy for facilitating specificity and sensitivity of nucleic acid detection, amplification, and gene cloning.

中文翻译：

脱氧核糖核酸聚合反应灵敏核酸检测的特异性增强

DNA聚合的特异性在DNA复制和遗传信息存储中起关键作用。同样，生物技术应用，例如核酸检测，DNA扩增和基因克隆，在由DNA聚合酶催化的DNA合成中要求高度特异性。但是，DNA聚合反应中的错误（例如错误掺入和错误引发）会严重损害特异性。在本文中，我们报告了我们的发现，即通过硫代磷酸酯dNTP减弱DNA聚合酶动力学，可以大大提高DNA酶促合成的特异性（最高100倍）。这种特异性增强可实现便捷，灵敏的核酸检测，聚合，PCR和复杂系统（例如人cDNA和基因组DNA）的基因克隆。进一步，我们发现特异性增强为检测核酸（例如COVID-19病毒RNA）提供了更高的灵敏度（提高了50倍）。我们的发现揭示了一种促进核酸检测，扩增和基因克隆的特异性和敏感性的简便方法。

更新日期：2020-12-15

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