<br>AMPLON：使用核酸的多臂引物和环优化扩增 DNA,Advanced Materials

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AMPLON：使用核酸的多臂引物和环优化扩增 DNA
Advanced Materials ( IF 27.4 ) Pub Date : 2024-04-24 , DOI: 10.1002/adma.202311634
Mert Tunca Doganay ₁ , Ethan Roman ₁ , Andrea M Hujer _{1,

2} , Robert A Bonomo _{1,

2,

3} , Steven G Deeks ₄ , Daniel R Kuritzkes _{5,

6} , Mohamed S Draz _{1,

3,

7,

8}

Affiliation

核酸扩增是生物技术和分子诊断的基石，其应用（尤其是等温方法）激增，增加了对先进和精确设计方法的需求。本文提出了一种使用核酸多臂引物和环优化（AMPLON）扩增 DNA 的新方法。AMPLON 依赖于一种新型聚合物材料和独特的多臂聚乙二醇-DNA 引物，可在等温条件下进行高效的 DNA 扩增。每个臂都携带单链 DNA，与目标 DNA 的正义或反义序列互补。扩增反应从反义臂与靶 DNA 结合开始，形成一个模板，用于携带正义臂通过连续的 DNA 环和解环步骤指导多臂大 DNA 扩增子合成。使用人类免疫缺陷病毒 1 型（HIV-1）作为模型临床靶点，AMPLON 表现出高灵敏度，可检测低至 100 拷贝 ^mL-1 的靶点浓度。与使用敏感引物的定量实时聚合酶链反应检测相比，AMPLON 能够可靠地鉴定血浆样品（n = 20）中的 HIV-1 RNA，一致性高达 95%。AMPLON 能够在 30 分钟内实现高度特异性和灵敏的靶标扩增，在改变核酸研究领域并释放医学和生物技术新的可能性方面具有巨大潜力。

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AMPLON: Amplifying DNA with Multiarm Priming and Looping Optimization of Nucleic Acid

Nucleic acid amplification, the bedrock of biotechnology and molecular diagnostics, surges in applications—especially isothermal approaches—heightening the demand for advanced and precisely engineered methods. Here, a novel approach for amplifying DNA with multiarm priming and looping optimization of nucleic acid (AMPLON) is presented. AMPLON relies on a novel polymeric material with unique set of multiarm polyethylene glycol–DNA primers for efficient DNA amplification under isothermal conditions. Each arm carries single-stranded DNA complementing the sense or antisense sequence of the target DNA. The amplification reaction begins with antisense arms binding to the target DNA, forming a template for sense-carrying arms to direct multiarm large DNA amplicon synthesis through successive DNA looping and unlooping steps. Using human immunodeficiency virus type 1 (HIV-1) as a model clinical target, AMPLON exhibits high sensitivity, detecting target concentrations as low as 100 copies mL⁻¹. Compared to a quantitative real-time polymerase chain reaction assay using sensitive primers, AMPLON reliably identifies HIV-1 RNA in plasma samples (n = 20) with a significant agreement rate of 95%. With its ability to achieve highly specific and sensitive target amplification within 30 min, AMPLON holds immense potential to transform the field of nucleic acid research and unleashing new possibilities in medicine and biotechnology.

更新日期：2024-04-24

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