Temperature-Modulation Categorization of Argonaute-Based Bioreactions Coupled with Amplification Methods for Nucleic Acid Detection

Argonaute (Ago), a programmable nuclease, has recently emerged as a promising tool for nucleic acid detection. To achieve higher sensitivity, a nucleic acid amplification step is often introduced into the Ago-based detection system, leading the process suffered from the issues of temperature compatibility and device complexity. To address these limitations, integrating multiple reactions within a portable temperature control device shows significant potential to streamline the development and practical application of these systems. However, there are few systematic studies guiding the design of the integrated Ago-based detection platforms. Here, this review summarizes the representative Ago-based bioreactions combined with different amplification methods. We begin with an overview of recent advances in Ago protein research, followed by a classification of the existing detection platforms based on their temperature regulation mechanisms: multi-temperature reactions and isothermal reactions. Additionally, the enzymatic properties of the widely used Ago proteins are also discussed in this review and their working temperatures are highlighted when integrated into a specific detection process. Finally, we explore the future prospects and challenges for integrated Ago-based detection systems, aiming to offer readers a comprehensive understanding of these bioreactions' unique characteristics, potential applications, and design principles for next-generation nucleic acid detection devices.