Advancements and strategies in simultaneous multi-target detection for analyte multiplexing

The simultaneous detection of multiple biomarkers has become increasingly important for healthcare diagnostics, food quality control, and environmental monitoring. Real-world samples are often complex, containing multiple analytes that may interact, leading to cross-reactivity and complicating reliable detection. In response, significant advances have been made in developing multiplexed analytical platforms that enable accurate, efficient, and early detection of disease markers or contaminants. This review systematically analyzes literature published from 2018 to mid-2025, following PRISMA guidelines, to evaluate strategies, technologies, and applications in multiplexed analyte detection platforms. It critically reviews various approaches, including optical, electrochemical, and molecular methods, all of which aim to detect multiple analytes simultaneously in one or more samples. This study also explores a broad range of applications in which multi-target detection is valuable, including real-time monitoring of environmental contaminants and clinical diagnostic settings, where early disease detection is vital. In conclusion, this work highlights current research directions in multi-analyte detection, its limitations, and potential future advancements. • Reviews techniques enabling multiplex biosensing across diverse platforms. • Summarizes recent papers on optical, electrochemical, and molecular sensors. • Outlines applications in healthcare, food safety, and environmental monitoring. • Discusses current challenges and prospects for point-of-care multiplex biosensors.