School of Environmental Science and Technology, Key Laboratory of Industrial Ecology and Environmental Engineering (Ministry of Education), Dalian University of Technology, Dalian, 116024, China; Dalian POCT Laboratory, Dalian, 116024, China.
School of Chemistry, The University of New South Wales, Sydney, New South Wales, 2052, Australia.
Biosens Bioelectron. 2024 Nov 15;264:116687. doi: 10.1016/j.bios.2024.116687. Epub 2024 Aug 20.
Uracil-DNA glycosylase (UDG), an enzyme for repairing uracil-containing DNA damage, is crucial for maintaining genomic stability. Simple and fast quantification of UDG activity is essential for biological assay and clinical diagnosis, since its aberrant level is associated with DNA damage and various diseases. Herein, we developed a fully integrated "sample in-signal out" distance-based paper analytical device (dPAD) for visual quantification of UDG using a flow-controlled uracil-rich DNA hydrogel (URDH). The uracil base sites contained in the DNA hydrogel are mis-incorporated with dUTP by rolling circle amplification (RCA), which simplifies the preparation process of the functionalized hydrogel. In the presence of UDG, the uracil in URDH can be recognized and removed to induce the permeability change of URDH, resulting in the visible distance signal along the paper channel. Using dPAD, as low as 6.4 × 10 U/mL of UDG (within 80 min) is visually identified without any instruments and complicated operations. This integrated dPAD is advantageous for its simplicity, cost effectiveness, and ease of use. We envision that it has the great potential for point-of-care testing (POCT) in DNA damage testing, personalized healthcare assessment, and biomedical applications.
尿嘧啶-DNA 糖基化酶（UDG）是一种修复含尿嘧啶 DNA 损伤的酶，对于维持基因组稳定性至关重要。简单快速地定量 UDG 活性对于生物测定和临床诊断至关重要，因为其异常水平与 DNA 损伤和各种疾病有关。在此，我们开发了一种完全集成的“样品进信号出”基于距离的纸分析装置（dPAD），用于使用流动控制的富含尿嘧啶的 DNA 水凝胶（URDH）可视化定量 UDG。DNA 水凝胶中含有的尿嘧啶碱基位点通过滚环扩增（RCA）与 dUTP 错误掺入，简化了功能化水凝胶的制备过程。在 UDG 的存在下，URDH 中的尿嘧啶可以被识别并去除，从而导致 URDH 的渗透性变化，从而在纸通道上产生可见的距离信号。使用 dPAD，即使在没有任何仪器和复杂操作的情况下，也可以在 80 分钟内以低至 6.4×10 U/mL 的 UDG 进行可视化识别。这种集成的 dPAD 具有简单、经济高效和易于使用的优点。我们设想它在 DNA 损伤检测、个性化医疗保健评估和生物医学应用中的即时检测（POCT）方面具有巨大的潜力。
