Biointerfaces Institute, McMaster University, 1280 Main Street West, Hamilton, ON, L8S 4K1, Canada.
Division of Respirology, McMaster University, and, Firestone Institute of Respiratory Health at St. Joseph's Health Care, Hamilton, ON, L8N 4A6, Canada.
Chembiochem. 2024 Jun 3;25(11):e202400085. doi: 10.1002/cbic.202400085. Epub 2024 Apr 29.
Over the last three decades, significant advancements have been made in the development of biosensors and bioassays that use RNA-cleaving DNAzymes (RCDs) as molecular recognition elements. While early examples of RCDs were primarily responsive to metal ions, the past decade has seen numerous RCDs reported for more clinically relevant targets such as bacteria, cancer cells, small metabolites, and protein biomarkers. Over the past 5 years several RCD-based biosensors have also been evaluated using either spiked biological matrixes or patient samples, including blood, serum, saliva, nasal mucus, sputum, urine, and faeces, which is a critical step toward regulatory approval and commercialization of such sensors. In this review, an overview of the methods used to generate RCDs and the properties of key RCDs that have been utilized for in vitro testing is first provided. Examples of RCD-based assays and sensors that have been used to test either spiked biological samples or patient samples are then presented, highlighting assay performance in different biological matrixes. A summary of current prospects and challenges for development of in vitro diagnostic tests incorporating RCDs and an overview of future directions of the field is also provided.
在过去的三十年中，使用 RNA 切割 DNA 酶 (RCD) 作为分子识别元件的生物传感器和生物测定法取得了重大进展。虽然早期的 RCD 主要对金属离子有反应，但在过去的十年中，已经有许多 RCD 被报道用于更具临床相关性的靶标，如细菌、癌细胞、小代谢物和蛋白质生物标志物。在过去的 5 年中，已经使用添加了生物基质或患者样本（包括血液、血清、唾液、鼻粘液、痰、尿液和粪便）的基于 RCD 的生物传感器进行了评估，这是朝着此类传感器的监管批准和商业化迈出的关键一步。在这篇综述中，首先提供了用于生成 RCD 的方法概述，以及已用于体外测试的关键 RCD 的特性。然后介绍了基于 RCD 的测定法和传感器的示例，这些示例用于测试添加的生物样本或患者样本，突出了在不同生物基质中的测定性能。还总结了包含 RCD 的体外诊断测试的当前前景和挑战，并概述了该领域的未来方向。
