| Summary: | Abstract Nanopore-based sensing has emerged as a transformative approach for biomarker detection, offering label-free, single-molecule analysis with exceptional sensitivity and specificity. By monitoring ionic current modulations as individual molecules traverse biological or solid-state nanopores, these platforms provide direct insights into molecular size, structure, charge, and interactions. Over the past five years, advances in nanopore fabrication, surface engineering, and signal interpretation have expanded their analytical scope, from nucleic acids and proteins to peptides and small metabolites, while enabling operation in complex biological matrices. This review focuses on recent advancements (from last 5 years) that enhance detection performance and specificity through recognition strategies, engineered pore chemistries, and data analysis. Applications are discussed across key disease areas, including cancer, cardiovascular, neurological, metabolic, and infectious diseases, emphasizing early detection, multiplexed measurements, clinical adaptability and translational diagnostics, highlighting progress toward portable, point-of-care systems. Collectively, these developments underscore the potential of nanopore biosensing to bridge fundamental research and real-world diagnostics, paving the way for rapid, sensitive, and accessible health monitoring tools in precision medicine. Graphical Abstract Schematic illustration of sensing of various health biomarkers (protein, peptides, miRNA, small molecules etc.) through nanopores.
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