Advances in Nucleic Acid-Based Amplification Strategies for Dual-Mode Biosensing of Pathogenic Bacteria: From Molecular Mechanisms to Integrated Analytical Systems.

Abstract

Rapid, accurate, and reliable detection of pathogenic bacteria remains a critical need in clinical, food, and environmental monitoring. In today's context, nucleic acid amplification-mediated biosensing have emerged as a prominent approach to improve detection sensitivity, whereas dual-mode signal readout approaches have more enhanced analytical robustness and reliability. This review outlines recent advances in nucleic acid signal amplification strategies, including enzyme-based methods like LAMP, RPA, and RCA, as well as enzyme-free approaches like HCR, CHA, and EDR. Special attention is given to incorporating these amplification methods into dual-mode biosensing systems that combine both optical and electrochemical transduction mechanisms. This integration enables complementary signal generation and improves detection accuracy by reducing false-positive and false-negative results. This study critically examines the advancement of nucleic acid signal amplification strategies (NASAS)-mediated dual-mode sensing systems for detecting major pathogenic bacteria, including Escherichia coli, Salmonella, Listeria monocytogenes, Staphylococcus aureus, and Vibrio species, focusing on selectivity, sensitivity, assay design, and real-sample applicability. Finally, the review highlights present challenges related to system integration, standardization, and point-of-care applications. Additionally, it outlines potential future directions for rendering nucleic acid amplification-based dual-mode probes into practical diagnostic devices. Overall, this study affords a comprehensive synthesis of emerging approaches and design mechanisms for next-generation diagnostic scaffold for pathogen analysis.