Biointerfaces Institute, McMaster University, 1280 Main Street West, Hamilton, ON, L8S 4O3, Canada.
Department of Biochemistry and Biomedical Sciences, McMaster University, 1280 Main Street West, Hamilton, ON, L8S 4K1, Canada.
Chem Soc Rev. 2022 Oct 31;51(21):9009-9067. doi: 10.1039/d2cs00613h.
Functional nucleic acids (FNAs), including DNA aptamers and DNAzymes, are finding increasing use as molecular recognition elements for point-of-care (POC) assays and sensors. An ongoing challenge in the development of FNA-based POC sensors is the ability to achieve detection of low levels of analyte without compromising assay time and ease of use. Rolling circle amplification (RCA) is a leading nucleic acid (NA) isothermal amplification method which can be coupled with FNAs for the ultrasensitive detection of non-NA targets. Herein we examine the key considerations required when designing FNA-coupled biosensors utilizing RCA. Specifically, we describe methods for using FNAs as inputs to regulate RCA, various modes of RCA amplification, and methods to detect the output of the RCA reaction, along with how these can be combined to allow detection of non-NA targets. Recent progress on development of portable optical and electrochemical POC devices that incorporate RCA is then described, followed by a summary of key challenges and opportunities in the field.
功能性核酸（FNAs），包括 DNA 适体和 DNA 酶，作为分子识别元件，在即时检测（POC）分析和传感器中得到了越来越多的应用。在基于 FNA 的 POC 传感器的开发中，一个持续存在的挑战是在不影响分析时间和易用性的情况下，实现对低水平分析物的检测。滚环扩增（RCA）是一种领先的核酸（NA）等温扩增方法，可与 FNAs 结合，用于非 NA 靶标的超灵敏检测。本文研究了在设计利用 RCA 的 FNA 偶联生物传感器时所需的关键考虑因素。具体来说，我们描述了将 FNAs 用作调节 RCA 的输入、各种 RCA 扩增模式以及检测 RCA 反应输出的方法，以及如何将它们结合起来以允许检测非 NA 靶标。然后描述了在开发包含 RCA 的便携式光学和电化学 POC 设备方面的最新进展，最后总结了该领域的关键挑战和机遇。
