Document Type
Article
Publication Date
4-24-2024
Manuscript Version
vor
Abstract
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.
Keywords
branched polyethylene glycol, HIV-1, isothermal amplification, multiarm primer, nucleic acid amplification
Publication Title
Advanced Materials
Grant
P30 DA054557
Rights
© 2024 The Authors. Advanced Materials published by Wiley-VCH GmbH. This is an open access article under the terms of the Creative Commons Attribution-NonCommercial-NoDerivs License, which permits use and distribution in any medium, provided the original work is properly cited, the use is non-commercial and no modifications or adaptations are made.
Creative Commons License
This work is licensed under a Creative Commons Attribution-NonCommercial-No Derivative Works 4.0 International License.
Recommended Citation
M. T. Doganay, E. Roman, A. M. Hujer, R. A. Bonomo, S. G. Deeks, D. R. Kuritzkes, M. S. Draz, AMPLON: Amplifying DNA with Multiarm Priming and Looping Optimization of Nucleic Acid. Adv. Mater. 2024, 36, 2311634. https://doi.org/10.1002/adma.202311634