Biosensors-Basel最新文献

Single-molecule sequencing technology, a novel method for gene sequencing, utilizes nano-sized materials to detect electrical and fluorescent signals. Compared to traditional Sanger sequencing and next-generation sequencing technologies, it offers significant advantages, including ultra-long read lengths, rapid sequencing, and the absence of amplification steps, making it widely applicable across various fields. By examining the development and components of single-molecule sequencing technology, it becomes clear that its unique characteristics provide new opportunities for advancing metrological traceability. Notably, its direct detection capabilities offer a novel approach to nucleic acid metrology. This paper provides a detailed overview of library construction, signal generation and detection, and data analysis methods in single-molecule sequencing and discusses its implications for nucleic acid metrology.

Recently, gold nanoclusters (AuNCs) have been widely used in biological applications due to their ultrasmall size, ranging within a few nanometers; large specific surface area; easy functionalization; unique fluorescence properties; and excellent conductivity. However, because they are unstable in solution, AuNCs require stabilization by using ligands such as dendrimers, peptides, DNA, and proteins. As a result, the properties of AuNCs and their formation are determined by the ligand, so the selection of the ligand is important. Of the many ligands implemented, enzyme-stabilized gold nanoclusters (enzyme-AuNCs) have attracted increasing attention for biosensor applications because of the excellent optical/electrochemical properties of AuNCs and the highly target-specific reactions of enzymes. In this review, we explore how enzyme-AuNCs are prepared, their properties, and the various types of enzyme-AuNC-based biosensors that use optical and electrochemical detection techniques. Finally, we discuss the current challenges and prospects of enzyme-AuNCs in biosensing applications. We expect this review to provide interdisciplinary knowledge about the application of enzyme-AuNC-based materials within the biomedical and environmental fields.

This study presents a novel microspectrometer-integrated microfluidic system for real-time protein concentration monitoring. The device employs electrokinetic principles for efficient protein preconcentration in a PDMS and Nafion film channel. Using FITC-labeled BSA as a model protein, the system demonstrated a linear correlation between protein concentration and absorbance at 491 nm. Notably, it achieved a 833-fold concentration increase from an initial 10 nM within 20 min. The compact microspectrometer system offers enhanced accuracy and sensitivity compared to traditional fluorescence microscopy methods. This innovation presents a promising solution for portable and point-of-care diagnostic applications, facilitating timely disease detection and monitoring. The findings highlight the potential for this technology to advance protein analysis and biomarker discovery in clinical settings, potentially improving patient outcomes through enhanced diagnostic capabilities.

Recent studies have shown that lactate is a molecule that plays an indispensable role in various physiological cellular processes, such as energy metabolism and signal transductions related to immune and inflammatory processes. For these reasons, interest in its detection using biosensors for non-invasive analyses of sweat during sports activity and in clinical reasons assessments has increased. In this minireview, an in-depth study was carried out on biosensors that exploited using electrochemical methods and innovative nanomaterials for lactate detection in sweat. This detection of lactate by biosensors in the sweat method seems to be feasible and highly desirable. From this commentary analysis, we can conclude that the correlation between lactate concentrations in sweat and blood is not yet clear, and studies are needed to clarify some key issues essential for the future application of this technology.

β-parvalbumin (β-PV) is the primary fish allergen responsible for most allergic reactions in individuals sensitive to fish. To ensure food safety, a sandwich-based magnetic immunoassay was developed using maleimide-functionalized magnetic beads (NH-MBs). Specific anti-β-PV antibodies were immobilized on these MBs, and a screen-printed carbon electrode was employed as the electrochemical transducer. A linear concentration range from 10 to 1000 ng/mL, a limit of detection of 1.8 ng/mL, and a limit of quantification of 7.1 ng/mL were achieved. Nineteen commercial food samples were analyzed to assess the potential of the sensor for routine use in food quality control. Important factors such as protein source and food processing (e.g., boiling, grilling, and frying) and preservation (e.g., in oil, and vacuum) were evaluated. The validated results confer the usefulness of the assay for food quality control.

Peroxynitrite (ONOO^-) plays an important role in many physiological and pathological processes. Excessive ONOO^- in cells leads to oxidative stress and inflammation. However, precise monitoring of ONOO^- levels in specific organelles (e.g., mitochondria) is still lacking and urgently needed. Herein, we rationally designed a mitochondria-targeted ratiometric fluorescent probe, MOBDP-I, for imaging of ONOO^- in the mitochondria of inflammatory cells and model mice. This probe, MOBDP-I, was synthesized by conjugating a BODIPY fluorophore to a mitochondria-targeting moiety-indole-salt group by a carbon-carbon double bond (C=C). In the presence of ONOO^-, the C=C bond between the BODIPY backbone and the indole-salt group was oxidized and broken, leading to an 18-fold enhancement of fluorescence at 510 nm, along with a significant fluorescence decrease at 596 nm. The ratiometric response property bestowed the probe with advantages in the precise quantification of ONOO^- in cells, thus allowing estimation of the extent of inflammation in living cells and mouse models of rheumatoid arthritis, peritonitis, and brain inflammation. MOBDP-I could act as an effective molecular tool to study the relationship between ONOO^- and the occurrence and development of inflammatory diseases.

Staphylococcus aureus (S. aureus) represents one of the most frequent worldwide causes of morbidity and mortality due to an infectious agent. It is a part of the infamous ESKAPE group, which is highly connected with increased rates of healthcare-associated infections and antimicrobial resistance. S. aureus can cause a large variety of diseases. Protein A (PrA) is a cell-wall-anchored protein of S. aureus with multiple key roles in colonization and pathogenesis and can be considered as a marker of S. aureus. The development of aptasensors, having an aptamer as a specific biorecognition element, increases selectivity, especially when working with complex matrices. The association with state-of-the-art materials, such as MXenes, can further improve the analytical performance. A competitive aptasensor configuration based on a ferrocene (Fc)-labeled cDNA hybridized (cDNA-Fc S13) on a specific aptamer (APT) for PrA in the presence of MXene nanosheets was designed for the indirect detection of S. aureus. The aptasensor displayed a linear range of 10-125 nM, an LOD of 3.33 nM, and a response time under 40 min. This configuration has been tested in real samples from volunteers diagnosed with S. aureus infections with satisfactory results, enabling the perspective to develop decentralized devices for the rapid detection of bacterial strains.

Vibrio parahaemolyticus (V. parahaemolyticus) is a significant concern, as it can cause severe infections and hemolytic trauma. Given its prevalence in seawater and coastal seafood, it poses a substantial risk as a foodborne pathogen. Biosensor-based detection technology has been continuously evolving, and toehold switches have emerged as a promising area within it, especially in the detection of RNA viruses. Here, we have developed a cell-free toehold switch sensor for V. parahaemolyticus detection. Traditional toehold switch detection methods usually use green fluorescent protein (GFP) or enzyme LacZ as the output signal, with an incubation time as long as 2 h, and are also mainly applied to the detection of RNA viruses. In this study, we introduced a novel, artificially designed luciferase (LuxSit-i) as an output signal and constructed toehold switches with two different output signals (sfGFP, LuxSit-i), aimed at reducing the incubation time of toehold switches. Moreover, to further improve the detection process, we separately utilize recombinase polymerase amplification (RPA) and nucleic acid sequence-based amplification (NASBA) to amplify dead and live bacterial suspensions for detection and attempt to distinguish between dead and live bacteria. This study provided a convenient, rapid, and accurate method for the on-site detection of V. parahaemolyticus, especially beneficial for resource-limited settings. By eliminating the requirement for specialized facilities and personnel, this system has the potential to be a valuable tool in improving public health responses, especially in developing regions.

Monitoring respiration rate (RR) is crucial in various healthcare settings, particularly during demanding (physical) activities where respiratory dynamics are critical indicators of health status. This study aimed to evaluate the accuracy of photoplethysmography (PPG)-based monitoring of RR during high-intensity interval training (HIIT) and its potential applications in healthcare. Between January and March 2024, healthy volunteers participated in a cycling HIIT session with increasing resistance levels. The RR measurements obtained using the PPG-based CardioWatch 287-2 (Corsano Health) were compared with an ECG patch-derived (Vivalink) reference. Subgroup analyses were conducted based on skin type and sex. A total of 35 participants contributed 1794 paired RR measurements. The PPG algorithm for RR monitoring showed an average root mean square (Arms) error of 2.13 breaths per minute (brpm), a bias of -0.09 brpm, and limits of agreement (LoA) from -4.28 to 4.09 brpm. Results were consistent across the different demographic subgroups. The CardioWatch 287-2 therefore demonstrated reliable RR monitoring during HIIT, supporting its potential use in healthcare settings for continuous, non-invasive respiratory monitoring, particularly in physical rehabilitation and chronic respiratory condition management.

Vascular endothelial growth factor (VEGF) is a critical angiogenesis biomarker associated with various pathological conditions, including cancer. This study leverages pre-biotinylated FcγRI interactions with IgG1-type monoclonal antibodies to develop a sensitive VEGF detection method. Utilizing surface plasmon resonance (SPR) technology, we characterized the binding dynamics of immobilized biotinylated FcγRI to an IgG1-type antibody, Bevacizumab (AVT), through kinetic studies and investigated suitable conditions for sensor surface regeneration. Subsequently, we characterized the binding of FcγRI-captured AVT to VEGF, calculating kinetic constants and binding affinity. A calibration curve was established to analyze the VEGF quantification capacity and accuracy of the biosensor, computing the limits of blank, detection, and quantification at a 95% confidence interval. Additionally, the specificity of the biosensor for VEGF over other protein analytes was assessed. This innovative biomimetic approach enabled FcγRI-mediated site-specific AVT capture, establishing a stable and reusable platform for detecting and accurately quantifying VEGF. The results indicate the effectiveness of the plasmonic sensor platform for VEGF detection, making it suitable for research applications and, potentially, clinical diagnostics. Utilizing FcγRI-IgG1 antibody binding, this study highlights the industrial and clinical value of advanced biosensing technologies, offering insights to enhance therapeutic monitoring and improve outcomes in anti-VEGF therapies.