Department of Chemical and Biomolecular Engineering (BK21+ Program), KAIST, 291 Daehak-ro, Yuseong-gu, Daejeon 34141, Republic of Korea.
Department of Biological Engineering, College of Engineering, Konkuk University, Seoul 05029, Republic of Korea.
Theranostics. 2020 Mar 15;10(10):4507-4514. doi: 10.7150/thno.41484. eCollection 2020.
Recently, personal glucose meter (PGM) has been utilized for the detection of non-glucose targets for point-of-care (POC) testing. Aimed at this goal, we herein developed a new PGM-based label-free read-out method for polymerase chain reaction (PCR) based on our novel finding that cerium oxide nanoparticles (CeO NPs) exhibit glucose oxidase-like activity comparable to the natural glucose oxidase enzyme. : In principle, DNA amplicons produced by PCR in the presence of target DNA electrostatically bind to CeO NPs, leading to their aggregation and reducing the efficiency for CeO NP-catalyzed glucose oxidation reaction. Thus, glucose is hardly oxidized to gluconic acid, resulting in the maintenance of initial high glucose level. On the contrary, in the absence of target DNA or presence of non-target DNA, DNA amplicons are not produced and glucose is effectively oxidized by the glucose oxidase-like activity of CeO NPs, leading to the significant reduction of glucose level. Finally, the resulting glucose level is simply measured by using PGM. : With this strategy, DNA amplicons were quantitatively examined within min, realizing ultrafast analysis of PCR results without any cumbersome and labor-intensive procedures. In addition, the target genomic DNA derived from () was sensitively determined down to copies with high selectivity. : Importantly, the use of PGM as a detection component enables its direct application in POC settings. Based on the meritorious features of PGM such as rapidity, simplicity, and cost-effectiveness, we expect that the devised system could serve as a core platform for the on-site read-out of PCR amplification.
最近,个人血糖仪 (PGM) 已被用于即时检测 (POC) 中非葡萄糖目标的检测。针对这一目标,我们开发了一种基于 PGM 的新型无标记读出方法,用于基于聚合酶链反应 (PCR) 的检测,其原理是基于我们的新发现,即氧化铈纳米粒子 (CeO NPs) 具有与天然葡萄糖氧化酶相当的葡萄糖氧化酶样活性。:原则上,存在靶 DNA 时 PCR 产生的 DNA 扩增子通过静电作用与 CeO NPs 结合,导致其聚集并降低 CeO NP 催化的葡萄糖氧化反应效率。因此,葡萄糖几乎不会被氧化为葡萄糖酸,导致初始高葡萄糖水平得以维持。相反,在不存在靶 DNA 或存在非靶 DNA 的情况下,不会产生 DNA 扩增子,并且葡萄糖会被 CeO NPs 的葡萄糖氧化酶样活性有效氧化,导致葡萄糖水平显著降低。最后,通过使用 PGM 简单测量产生的葡萄糖水平。:通过这种策略,在 min 内定量检测了 DNA 扩增子,实现了 PCR 结果的超快速分析,无需任何繁琐和劳动密集型的步骤。此外,还可以高度选择性地检测到来自 () 的靶基因组 DNA 低至 拷贝。:重要的是,将 PGM 用作检测组件可使其直接应用于 POC 环境。基于 PGM 的快速、简单和经济高效等优点,我们预计所设计的系统可以作为 PCR 扩增现场读出的核心平台。