Biosensors have become indispensable tools in clinical diagnostics, transforming how diseases are detected, monitored, and managed. In this review, we survey six key emerging platform classes: engineered biorecognition biosensors (CRISPR-Cas, aptamer, molecularly imprinted polymers); electrochemical and electrical biosensors (graphene/MXene/quantum-dot nanomaterial transducers, field-effect, electrochemical impedance spectroscopy, photoelectrochemical sensing); optical and plasmonic biosensors (SPR, LSPR, SERS); lateral flow and paper-based assays; artificial intelligence-integrated smart diagnostics; and lab-on-chip microfluidic systems. For each, we analyze the recognition chemistry, transduction strategy, and reported clinical performance. We then map these technologies onto high-impact clinical domains: infectious disease, oncology, cardiovascular medicine, diabetes and metabolic monitoring, critical care and sepsis, and neurodegenerative disease—highlighting representative and quantitative performance metrics and explicitly distinguishing analytical performance from clinical validation status using a six-level validation framework. Finally, we discuss the translational barriers (reproducibility, matrix complexity, manufacturing scalability, regulatory pathways, and biocompatibility) and the opportunities that health-economic evidence for microfluidic platforms, AI integration, and wearable and implantable form factors offer for closing the gap between analytical performance demonstrated in laboratory settings and real-world clinical adoption.
Semiconductor–noble metal hybrids have emerged as attractive surface-enhanced Raman scattering (SERS) platforms for pharmaceutical analysis because they integrate interfacial chemical interactions with plasmon-mediated electromagnetic enhancement. Herein, a semiconductor–noble metal hybrid SERS substrate was constructed by sputtering Ag nanoparticles (NPs) onto exfoliated MoS2 nanosheets (NSs) and was applied to the quantitative detection of ibrutinib. Within this hybrid architecture, accessible surfaces for analyte adsorption were provided by the exfoliated MoS2 NSs, while plasmonically coupled electromagnetic “hotspots” were generated by the Ag NPs, as further supported by finite-difference time-domain (FDTD) simulations. The optimized substrate enabled quantitative SERS detection of ibrutinib over 10−6–10−2 mg mL−1 (R2 = 0.991), with a detection limit of 3.73 × 10−8 mg mL−1 and a relative standard deviation of 5.81%. These results demonstrate the potential of Ag-decorated MoS2 NSs for sensitive SERS detection of ibrutinib in standard solutions.
DNA adenine methyltransferase (Dam MTase) is a key epigenetic biomarker linked to bacterial pathogenicity and tumor progression, holding great value for disease diagnosis and drug discovery. Conventional Dam MTase detection approaches suffer from complex workflows, long assay times, limited sensitivity and unsatisfactory anti-interference ability, severely hindering their real-world implementation. Herein, a facile fluorescent biosensor was constructed for rapid Dam MTase activity detection by integrating a hairpin DNA probe, endonuclease cleavage inhibition and isothermal cyclic amplification. In the absence of Dam MTase, Nt.AlwI specifically cleaves the engineered hairpin probe to trigger amplification and generate intense fluorescence. Conversely, Dam-induced DNA methylation prevents enzymatic cleavage, suppresses amplification and reduces fluorescence output, where fluorescence intensity varies with Dam MTase activity in a concentration-dependent fashion. Experimental validation confirms high specificity and favorable anti-interference performance of this sensor under complex biological conditions. It enables inhibitor screening and reliable measurement of endogenous Dam MTase activity in bacterial samples. With simple configuration, easy manipulation and low cost, the biosensor addresses major drawbacks of classic assays and provides a robust analytical platform for epigenetic studies, clinical bacterial testing and high-throughput targeted-inhibitor screening.
The morphology of the functional coating (thickness, density, homogeneity) determines the accuracy and reproducibility ofabel-free biosensor analysis, yet the quality of the coating is usually assessed indirectly, outside the conditions of the analysis. We describe an in situ method for functional assessment of the coating by the biosensor’s own response, based on surface waves in a one-dimensional photonic crystal with two-dimensional imaging. In one cycle of glutaraldehyde activation and protein immobilization, we determine sorption capacity, interchannel homogeneity, flow stability and surface density. At a manyfold excess of reactive centers, the saturated binding signal is insensitive to theocal density of functional groups. On a series of 3-aminopropyltriethoxysilane (APTES) coatings, the sorption capacity and homogeneity depend critically on the solvent composition, and the solution-front delay maps separate the coating nonuniformity from mass transfer. For the optimized coating, formed in a 40% solution of acetic acid in ethanol, where theow water content and acidity suppress the self-condensation of APTES, the surface concentration of bovine serum albumin reaches 1.98 mg/m2, a dense side-on monolayer with a coefficient of variation of 1.2%. The results are confirmed by IgG immobilization and streptavidin–biotin binding. The approach enables noninvasive control of biofunctionalization.
Low-intensity pulsed ultrasound (LIPUS) has antitumor potential, but its effects on tumor cell mechanics remain poorly understood. In this study, photonic crystal cellular force microscopy (PCCFM) enabled time-resolved mechanical measurements of colorectal cancer cells after LIPUS treatment. Serial PCCFM structural-color and corresponding bright-field images were acquired to reconstruct the vertical stress and substrate deformation in cell-associated regions; integration of the reconstructed stress over defined regions yielded quantitative collective and single-cell vertical force measurements LIPUS altered the time course of collective cellular force after seeding exhibiting distinct collective response patterns. Single-cell analysis further showed marked intercellular heterogeneity in force magnitude and temporal profiles. These mechanical changes were accompanied by altered F-actin organization, fewer peripheral protrusions, and reduced migration and invasion indices, indicating concordant changes in cellular force and motility-related structural and functional phenotypes. In vivo, xenografts derived from LIPUS-pretreated cells had reduced final xenograft volume and tumor weight. Piezo1 immunofluorescence and Ca2+ imaging suggested an association between mechanosensitive-channel signaling and the observed mechanical changes. Overall, PCCFM resolved temporal vertical force responses to LIPUS and revealed associations among LIPUS treatment, cellular mechanics, and motility- and growth-related phenotypes. Therefore, PCCFM provides a label-free, visual, and quantitative cellular-force sensing approach for assessing the dynamic responses of tumor cells to external mechanical stimulation.
