Darius Aw Kang Lie, Ling Neo Jia, Mahesh Uttamchandani
School of Chemical and Biomedical Engineering, Nanyang Technological University, 637459, Singapore.
Mol Biosyst. 2010 May;6(5):792-4. doi: 10.1039/c001923b. Epub 2010 Mar 19.
The split G-quadruplex DNAzyme has emerged as a valuable tool for visual DNA detection. Most reports on its use have however been constrained to proof-of-concept demonstrations using synthetic oligonucleotides. This is due to the inherent complexities of the assay, and the multiple components involved. Herein, we have overcome several of these challenges and have successfully integrated asymmetric PCR with the visual detection step, allowing enriched targets from complex samples to be conveniently detected. This workflow would enable the DNAzyme system to be applied for point-of-care DNA detection applications. We have established the platform herein as a simple and robust method to determine the presence of target DNA sequences post-PCR (as required for pathogen detection), without the need for gels or other apparatus. Nanomolar concentrations of amplified targets were detected using the workflow established, enabling the detection of salmonella and mycobacterium targets through a color change reaction, observable just by eye.
分裂型G-四链体DNA酶已成为一种用于可视化DNA检测的有价值工具。然而，大多数关于其应用的报告都局限于使用合成寡核苷酸的概念验证演示。这是由于该检测方法固有的复杂性以及所涉及的多个组件。在此，我们克服了其中的几个挑战，并成功地将不对称PCR与可视化检测步骤相结合，从而能够方便地检测复杂样品中富集的目标。这种工作流程将使DNA酶系统能够应用于即时护理DNA检测应用。我们在此建立了一个简单而稳健的平台，用于在PCR后确定目标DNA序列的存在（这是病原体检测所必需的），无需使用凝胶或其他仪器。使用所建立的工作流程检测到了纳摩尔浓度的扩增目标，通过肉眼可见的颜色变化反应能够检测沙门氏菌和分枝杆菌目标。
