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One-Pot Colorimetric Nucleic Acid Test Mediated by Silver Nanoparticles for DNA Extraction and Detection
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Rapid and Highly Sensitive Detection of Ricin in Biological Fluids Using Optical Modulation Biosensing
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Understanding the Mechanism of Bent DNA Amplifying Sensors Using All-Atom Molecular Dynamics Simulations
Journal Description
Biosensors
Biosensors
is an international, peer-reviewed, open access journal on the technology and science of biosensors published monthly online by MDPI.
- Open Access— free for readers, with article processing charges (APC) paid by authors or their institutions.
- High Visibility: indexed within Scopus, SCIE (Web of Science), PubMed, MEDLINE, PMC, Embase, CAPlus / SciFinder, Inspec, and other databases.
- Journal Rank: JCR - Q1 (Instruments and Instrumentation) / CiteScore - Q1 (Instrumentation)
- Rapid Publication: manuscripts are peer-reviewed and a first decision is provided to authors approximately 21.8 days after submission; acceptance to publication is undertaken in 2.8 days (median values for papers published in this journal in the first half of 2025).
- Recognition of Reviewers: reviewers who provide timely, thorough peer-review reports receive vouchers entitling them to a discount on the APC of their next publication in any MDPI journal, in appreciation of the work done.
Impact Factor:
5.6 (2024);
5-Year Impact Factor:
5.7 (2024)
Latest Articles
The Impact of Heat Stress on Dairy Cattle: Effects on Milk Quality, Rumination Behaviour, and Reticulorumen pH Response Using Machine Learning Models
Biosensors 2025, 15(9), 608; https://doi.org/10.3390/bios15090608 (registering DOI) - 15 Sep 2025
Abstract
Heat stress has a major impact on dairy cow health and productivity, especially during early lactation. Conventional heat stress monitoring methods frequently rely on single indicators, such as the temperature–humidity index (THI), which may miss subtle physiological and metabolic responses. This study presents
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Heat stress has a major impact on dairy cow health and productivity, especially during early lactation. Conventional heat stress monitoring methods frequently rely on single indicators, such as the temperature–humidity index (THI), which may miss subtle physiological and metabolic responses. This study presents a novel threshold-based classification framework that integrates biologically meaningful combinations of environmental, behavioural, and physiological variables to detect early-stage heat stress responses in dairy cows. Six composite heat stress conditions (C1–C6) were developed using real-time THI, milk temperature, reticulorumen pH, rumination time, milk lactose, and milk fat-to-protein ratio. The study applied and assessed five supervised machine learning models (Partial Least Squares Discriminant Analysis (PLS-DA), Support Vector Machine (SVM), Random Forest (RF0, Neural Network (NN), and an Ensemble approach) trained on daily datasets gathered from early-lactation dairy cows fitted with intraruminal boluses and monitored through milking parlour sensor systems. The dataset comprised approximately 36,000 matched records from 200 cows monitored over 60 days. The highest classification performance was observed for RF and NN models, particularly under C1 (THI > 73 and milk temperature > 38.6 °C) and C6 (THI > 74 and milk temperature > 38.7 °C), with AUC values exceeding 0.90. SHAP analysis revealed that milk temperature, THI, rumination time, and milk lactose were the most informative features across conditions. This integrative approach enhances precision livestock monitoring by enabling individualised heat stress risk classification well before clinical or production-level consequences emerge.
Full article
(This article belongs to the Special Issue Artificial Intelligence (AI) and Machine Learning (ML) in Biosensors: Innovation, Application, and Challenge)
Open AccessArticle
Gold Nanoparticle-Enhanced Recombinase Polymerase Amplification for Rapid Visual Detection of Mycobacterium tuberculosis
by
Sukanya Saikaew, Sirikwan Sangboonruang, Rodjana Pongsararuk, Prapaporn Srilohasin, Bordin Butr-Indr, Sorasak Intorasoot, Ponrut Phunpae, Chayada Sitthidet Tharinjaroen, Surachet Arunothong, Wutthichai Panyasit, Angkana Chaiprasert, Khajornsak Tragoolpua and Usanee Wattananandkul
Biosensors 2025, 15(9), 607; https://doi.org/10.3390/bios15090607 (registering DOI) - 15 Sep 2025
Abstract
Tuberculosis (TB) remains a major global health challenge, particularly in resource-limited settings where access to rapid and reliable diagnostics is limited. Conventional diagnostic methods, such as smear microscopy and culture, are either time-consuming or lack adequate sensitivity. This study optimized recombinase polymerase amplification
[...] Read more.
Tuberculosis (TB) remains a major global health challenge, particularly in resource-limited settings where access to rapid and reliable diagnostics is limited. Conventional diagnostic methods, such as smear microscopy and culture, are either time-consuming or lack adequate sensitivity. This study optimized recombinase polymerase amplification (RPA) using 16 primer combinations targeting IS6110 highly specific to the Mycobacterium tuberculosis complex (MTC). A novel naked-eye assay, TB-GoldDx, was developed by integrating RPA combined with gold nanoparticles (AuNPs), enabling equipment-free diagnostics. TB-GoldDx demonstrated a detection limit of 0.001 ng of MTB H37Rv DNA (~210 bacilli) per 25 µL reaction. Among 100 bacterial strains, it achieved 95.83% sensitivity and 100% specificity among 100 bacterial strains, comprising 72 MTB isolates and 28 nontuberculous bacterial species. In 140 sputum samples, the assay showed 81.43% sensitivity and 58.57% specificity versus acid-fast bacilli (AFB) smear microscopy, with sensitivity improving to 95.45% in high-load AFB 3+ specimens. Compared to a commercial line probe assay (LPA), TB-GoldDx exhibited slightly higher sensitivity (84.78% vs. 82.61%) but lower specificity (54.05% vs. 78.38%). Delivering rapid, visual results in under an hour, TB-GoldDx offers a low-cost, easily deployable solution for point-of-care tuberculosis detection, especially in underserved regions, reinforcing global End TB efforts.
Full article
(This article belongs to the Section Biosensors and Healthcare)
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Open AccessArticle
Terahertz High-Sensitivity SPR Phase Biosensor Based on the Weyl Semimetals
by
Yu Xie, Zean Shen, Mengjiao Ren, Mingming Zhang, Mingwei Guo and Leyong Jiang
Biosensors 2025, 15(9), 606; https://doi.org/10.3390/bios15090606 - 15 Sep 2025
Abstract
Optical biosensors play a crucial role in the field of biological detection by converting biological signals into optical signals for detection. Among them, Surface Plasmon Resonance (SPR) optical biosensors have become a research hotspot in this field due to their significant advantage of
[...] Read more.
Optical biosensors play a crucial role in the field of biological detection by converting biological signals into optical signals for detection. Among them, Surface Plasmon Resonance (SPR) optical biosensors have become a research hotspot in this field due to their significant advantage of high sensitivity. Weyl Semimetals (WSMs), as a type of three-dimensional topological material with unique electronic structures and other properties, exhibit potential applications in the field of SPR sensing. Against this background, we designed a terahertz (THz) high-sensitivity SPR phase biosensor with a KR structure based on WSMs. When applied in gas sensing scenarios, the phase detection sensitivity of this sensor can reach 22,402 /RIU, showing a significant improvement compared to traditional SPR biosensors. Moreover, we found that the Weyl node separation distance and twist angle of WSMs have obvious effects on sensitivity regulation. Additionally, we optimized the sensitivity and structural parameters of this structure using a neural network-based deep learning algorithm. We expect that this proposed scheme can provide a feasible reference for the field of biological sensing.
Full article
(This article belongs to the Special Issue New Progress in Optical Fiber-Based Biosensors—2nd Edition)
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Open AccessCommunication
An Aptamer-Based gFET-Sensor for Specific Quantification of Gene Therapeutic Human Adenovirus Type 5
by
Runliu Li, Ann-Kathrin Kissmann, Hu Xing, Roger Hasler, Christoph Kleber, Wolfgang Knoll, Hannes Schmietendorf, Tatjana Engler, Lea Krutzke, Stefan Kochanek and Frank Rosenau
Biosensors 2025, 15(9), 605; https://doi.org/10.3390/bios15090605 - 14 Sep 2025
Abstract
The combination of rGO-FETs (reduced Graphene Oxide Field-Effect Transistors) and DNA-oligonucleotide aptamers to sense analytes has been shown to be a promising technological approach, achieving high sensitivity and selectivity. With human adenovirus type 5 (HAdV-5) particles as the target, we here demonstrate the
[...] Read more.
The combination of rGO-FETs (reduced Graphene Oxide Field-Effect Transistors) and DNA-oligonucleotide aptamers to sense analytes has been shown to be a promising technological approach, achieving high sensitivity and selectivity. With human adenovirus type 5 (HAdV-5) particles as the target, we here demonstrate the application of the aptamer/FET combination for detection of this medically and biotechnologically relevant viral vector. A focused anti-HAdV-5 aptamer library was evolved in a nine-round SELEX process, allowing for the specific fluorescent labeling of HAdV-5 and related subtypes. Moreover, this library was already sufficient to serve as the binding entity on a gFET sensor for sensitive quantification of the virus particles. Adenoviruses have been widely used as gene delivery vectors for gene therapy and genetic vaccination. The use of adenoviral vectors within the vaccination campaign against COVID-19 emphasized the need for robust biotechnological production processes, which additionally require sensitive product formation monitoring. We believe that these type of gFET-based aptasensors can serve as the technological monitoring basis in virus production processes in the near future.
Full article
(This article belongs to the Special Issue Transistor-Based Biosensors and Their Applications)
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Open AccessArticle
Construction of an Electrochemical Impedance Spectroscopy Matching Method Based on Adaptive Multi-Error Driving and Application Testing for Biofilm Impedance Verification
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Hanyang Bao, Fan Yu, Peiyan Dai, Boyu Guo and Ying Xu
Biosensors 2025, 15(9), 604; https://doi.org/10.3390/bios15090604 - 12 Sep 2025
Abstract
Electrochemical impedance spectroscopy (EIS) is a technique used to analyze the kinetics and interfacial processes of electrochemical systems. The selection of an appropriate equivalent circuit model for EIS interpretation was traditionally reliant on expert experience, rendering the process subjective and prone to error.
[...] Read more.
Electrochemical impedance spectroscopy (EIS) is a technique used to analyze the kinetics and interfacial processes of electrochemical systems. The selection of an appropriate equivalent circuit model for EIS interpretation was traditionally reliant on expert experience, rendering the process subjective and prone to error. To address these limitations, an automated framework for both model selection and parameter estimation was proposed. The methodology was structured such that initial model screening was performed by a global heuristic search algorithm, adaptive optimization was guided by an integrated XGBoost-based error feedback mechanism, and precise parameter estimation was achieved using a Differential Evolution–Levenberg–Marquardt (DE-LM) algorithm. When evaluated on a purpose-built dataset comprising 4.8 × 105 spectra across diverse circuit and biofilm scenarios, a model classification accuracy of 96.32% was achieved, and a 72.3% reduction in parameter estimation error was recorded. The practical utility of the method was validated through the quantitative analysis of bovine serum albumin–Clenbuterol hydrochloride (BSA-CLB), wherein an accuracy of 95.2% was demonstrated and a strong linear correlation with target concentration (R2 = 0.999) was found. Through this approach, the limitations of traditional black-box models were mitigated by resolving the physical meaning of parameters. Consequently, the automated and quantitative monitoring of processes such as biofilm formation was facilitated, enabling the efficient evaluation of antimicrobial drugs or anti-fouling coatings.
Full article
(This article belongs to the Section Biosensor and Bioelectronic Devices)
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Open AccessReview
From Biomarkers to Biosensors: Modern Approaches for the Detection of Matrix Metalloproteinases (MMPs)
by
Raja Chinnappan, Lohit Ramachandran, Isha Uttam, Marimuthu Citartan, Nidambur Vasudev Ballal and Naresh Kumar Mani
Biosensors 2025, 15(9), 603; https://doi.org/10.3390/bios15090603 - 12 Sep 2025
Abstract
Matrix metalloproteinases (MMPs) are a class of extracellular Zn2+ peptidases involved in various physiological and pathological processes. These enzymes serve as excellent biomarkers for diagnosing various diseases, including cancer and periodontitis, to name a few. MMP levels also serve as a prognostic
[...] Read more.
Matrix metalloproteinases (MMPs) are a class of extracellular Zn2+ peptidases involved in various physiological and pathological processes. These enzymes serve as excellent biomarkers for diagnosing various diseases, including cancer and periodontitis, to name a few. MMP levels also serve as a prognostic marker, which helps determine how much the disease has progressed. However, the current methods used to detect MMPs need a large sample volume, carry a high cost, and are not widely accessible to the public due to these challenges. Biosensing techniques tackle these problems by providing an efficient, cost-effective sensor with great sensitivity. This review provides a comprehensive overview of the latest developments and advancements in detecting MMPs using biosensors that employ various detection mechanisms such as electrochemical, colorimetric, and fluorescence methods. Furthermore, we have discussed the challenges and prospects of using MMPs as diagnostic tools.
Full article
(This article belongs to the Special Issue Lighting Up Single-Molecule Biosensors and Bioimaging: Now and the Decade to Come)
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Open AccessArticle
Self-Adaptive Polymer Fabry–Pérot Thermometer for High-Sensitivity and Wide-Linear-Range Sensing
by
Yifan Cheng, Maolin Yu, Junjie Liu, Yingling Tan and Jinhui Chen
Biosensors 2025, 15(9), 602; https://doi.org/10.3390/bios15090602 - 12 Sep 2025
Abstract
Fiber-optic temperature sensors with advantages such as simplicity, low cost, and high sensitivity have attracted increasing attention. In this work, we propose a self-adaptive polymer Fabry–Pérot interferometer (PFPI) sensor for ultrasensitive and wide-linear-range thermal sensing. This design achieves a temperature sensitivity of 0.95
[...] Read more.
Fiber-optic temperature sensors with advantages such as simplicity, low cost, and high sensitivity have attracted increasing attention. In this work, we propose a self-adaptive polymer Fabry–Pérot interferometer (PFPI) sensor for ultrasensitive and wide-linear-range thermal sensing. This design achieves a temperature sensitivity of 0.95 nm/°C, representing an enhancement of two orders of magnitude compared to conventional fiber Bragg gratings. To address the challenge of spectral shifts exceeding the free spectral range due to the high sensitivity, a local cross-correlation algorithm is introduced for accurate wavelength tracking. We demonstrate ultrahigh-resolution (0.025 °C) scanning thermal field imaging and sensitive human physiological monitoring, including precise body temperature and respiratory rate detection. These results highlight the dual capability of our PFPI sensor for both microscopic thermal mapping and non-invasive healthcare applications.
Full article
(This article belongs to the Special Issue Polymers-Based Biosensors and Bioelectronics: Designs and Applications)
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Open AccessArticle
PEI-Fe3O4/PTA-AuNPs Hybrid System for Rapid DNA Extraction and Colorimetric LAMP Detection of E. faecium
by
Muniyandi Maruthupandi, Haang Seok Choi and Nae Yoon Lee
Biosensors 2025, 15(9), 601; https://doi.org/10.3390/bios15090601 - 12 Sep 2025
Abstract
This study introduces a novel nucleic acid testing (NAT) protocol that integrates rapid deoxyribonucleic acid (DNA) extraction, isothermal amplification, and visual detection to enable efficient analysis of opportunistic pathogens. Polyethylenimine-functionalized iron oxide (PEI-Fe3O4) nanoparticles were prepared by combining PEI,
[...] Read more.
This study introduces a novel nucleic acid testing (NAT) protocol that integrates rapid deoxyribonucleic acid (DNA) extraction, isothermal amplification, and visual detection to enable efficient analysis of opportunistic pathogens. Polyethylenimine-functionalized iron oxide (PEI-Fe3O4) nanoparticles were prepared by combining PEI, acting as a stabilizing agent, with iron salt, which was utilized as the metal ion precursor by the ultrasonication-assisted co-precipitation method, and characterized for structural, optical, and magnetic properties. PEI-Fe3O4 exhibited cationic and anionic behavior in response to pH variations, enhancing adaptability for DNA binding and release. PEI-Fe3O4 enabled efficient extraction of E. faecium DNA within 10 min at 40 °C, yielding 17.4 ng/µL and achieving an extraction efficiency of ~59% compared to a commercial kit (29.5 ng/µL). The extracted DNA was efficiently amplified by loop-mediated isothermal amplification (LAMP) at 65 °C for 45 min. Pyrogallol-rich poly(tannic acid)-stabilized gold nanoparticles (PTA-AuNPs) served as colorimetric probes for direct visual detection of the DNA amplified using LAMP. The magnetic-nanogold (PEI-Fe3O4/PTA-AuNPs) hybrid system achieved a limit of quantification of 1 fg/µL. To facilitate field deployment, smartphone-based RGB analysis enabled quantitative and equipment-free readouts. Overall, the PEI-Fe3O4/PTA-AuNPs hybrid system used in NAT offers a rapid, cost-effective, and portable solution for DNA detection, making the system suitable for microbial monitoring.
Full article
(This article belongs to the Special Issue Aptamer-Based Sensing: Designs and Applications)
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Open AccessReview
Metal–Phenolic Networks for Sensing Applications
by
Ning Xia, Sirui Liang, Dehua Deng, Yong Chang and Xinyao Yi
Biosensors 2025, 15(9), 600; https://doi.org/10.3390/bios15090600 - 11 Sep 2025
Abstract
The preparation of new inorganic–organic hybrid materials is beneficial for the development of powerful sensing methods and technologies. Polyphenols, a type of organic molecule containing phenolic hydroxyl groups, are widely present in natural plants and have beneficial effects on human health. Metal ions
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The preparation of new inorganic–organic hybrid materials is beneficial for the development of powerful sensing methods and technologies. Polyphenols, a type of organic molecule containing phenolic hydroxyl groups, are widely present in natural plants and have beneficial effects on human health. Metal ions are ubiquitous in nature and play an important role in the development of inorganic–organic hybrid materials. Metal–phenolic networks (MPNs) are formed by the self-assembly of metal ions and polyphenols through dynamic coordination bonds. Due to their mild synthesis conditions, facilely engineered functionalities, and multiple modification strategies, MPNs have become potential platforms for sensing applications. Timely understanding of the function and application of MPNs in sensing fields will facilitate the development of novel chemical and biological sensors and devices. This article summarizes the typical preparation methods and excellent advantages of MPNs and focuses on their latest achievements in sensing applications. We highlight representative MPN-based sensing examples, including the direct detection of small molecules and biological species, immunoassays, bioimaging, and wearable devices. Finally, the prospects and future directions of MPNs in sensing fields are addressed.
Full article
(This article belongs to the Special Issue Biosensors Based on Self-Assembly and Molecular Recognition—2nd Edition)
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Open AccessArticle
Foreign Body Reaction to Neural Implants: A Comparative Study of Polymer Toxicity and Tissue Response
by
Ciara Makievskaya, Anna Brezgunova, Nadezda Andrianova, Evgeny Kelm, Maria Solovyova, Gelena Naumova, Alina Zeinalova, Olga Gancharova, Tatiana Bushkova, Daniil Kozlov, Valery Putlayev, Pavel Evdokimov, Alexander Petrov, Mikhail Lebedev, Egor Plotnikov and Vasily Popkov
Biosensors 2025, 15(9), 599; https://doi.org/10.3390/bios15090599 - 11 Sep 2025
Abstract
This study investigated the toxicity of ten polymer materials intended for the development of invasive neural interfaces improving the treatment of neurological diseases. Most of the materials for neural implants can cause traumatization of the surrounding tissue, inflammation, and foreign body reaction. In
[...] Read more.
This study investigated the toxicity of ten polymer materials intended for the development of invasive neural interfaces improving the treatment of neurological diseases. Most of the materials for neural implants can cause traumatization of the surrounding tissue, inflammation, and foreign body reaction. In this study, in vitro and in vivo toxicity assessment was performed for nylon 618 (NY), polycaprolactone (PCL), polyethylene glycol diacrylate (PEGDA), polydimethylsiloxane (PDMS), polyethylene terephthalate (PET), polylactide (PLA), thermoplastic polyurethane (TPU), polypropylene (PP), polyethylene terephthalate glycol (PET-G), and polyimide (PI). The biocompatibility of these ten materials was assessed based on cell adhesion, growth and cytotoxicity on neural (PC-12) and fibroblast (NRK-49F) cultures. Furthermore, brain tissue responses to the implanted phantom scaffolds were analyzed in rats. According to these measurements, PI showed the highest compatibility for both cell types. PEGDA exhibited cytotoxic effects, low cell adhesion and the strongest foreign body reaction, including fibrosis and multinucleated cell formation. The other polymers showed lower pathological responses which makes them potentially usable for neural interfacing. We conclude that PEGDA appears to be unsuitable for long-term use due to adverse tissue and cellular reactions, whereas PI, PLA, PDMS and TPU hold promise as materials for safe and effective neural interface applications.
Full article
(This article belongs to the Special Issue Implantable, Wireless Biosensors and Biodevices for Neuroscience Research, 2nd Edition)
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Open AccessArticle
Evaluation of Fluorescence Detection Algorithms for Efficient ROI Setting in Low-Cost Real-Time PCR Systems
by
Seul-Bit-Na Koo, Ji-Soo Hwang, Chan-Young Park and Deuk-Ju Lee
Biosensors 2025, 15(9), 598; https://doi.org/10.3390/bios15090598 - 10 Sep 2025
Abstract
This study proposes a region of interest (ROI) setting method to improve the accuracy and efficiency of fluorescence detection in a compact real-time multiplex fluorescence PCR system. Conventional commercial real-time PCR systems are limited in point-of-care (POC) environments due to their high cost
[...] Read more.
This study proposes a region of interest (ROI) setting method to improve the accuracy and efficiency of fluorescence detection in a compact real-time multiplex fluorescence PCR system. Conventional commercial real-time PCR systems are limited in point-of-care (POC) environments due to their high cost and complex optical structures. To address this issue, we developed a low-cost, compact system using an open-platform camera and a Fresnel lens. However, in such a simply structured system, variations between the wells of the polymerase chain reaction (PCR) plate may affect the accuracy of fluorescence detection. In this study, after capturing images with a CMOS camera, we propose two ROI image processing algorithms. The proposed algorithms reliably extract fluorescence signals and compare ROI deviations caused by variations between wells to determine whether physical correction is necessary. To validate the system, we performed comparative analysis of real-time DNA amplification images and fluorescence dye images collected over multiple periods. Based on evaluations using manual detection as a reference, it was confirmed that even a simple algorithm can achieve stable fluorescence detection while minimizing ROI distortion. This study presents an efficient method for enhancing the accuracy of quantitative fluorescence analysis in small PCR systems and is expected to contribute to improving the performance of point-of-care diagnostics, thereby increasing accessibility to on-site diagnostics in the future.
Full article
(This article belongs to the Special Issue Recent Advances and Applications of Multiplexed Analysis and Multiplexed Nanobiosensors)
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Open AccessArticle
Simulation-Based Performance Assessment of Bulk Junctionless FET with Asymmetric Source/Drain for Ultrasensitive Detection of Biomolecules
by
Jeongmin Son, M. Meyyappan and Kihyun Kim
Biosensors 2025, 15(9), 597; https://doi.org/10.3390/bios15090597 - 10 Sep 2025
Abstract
Bio field-effect transistors (BioFETs) have attracted attention for their ability to rapidly detect physiological data with a simple structure. While conventional BioFETs offer high sensitivity, they often require reference electrodes or involve complex fabrication processes. A recently proposed bulk junctionless BioFET (Bulk JL-BioFET)
[...] Read more.
Bio field-effect transistors (BioFETs) have attracted attention for their ability to rapidly detect physiological data with a simple structure. While conventional BioFETs offer high sensitivity, they often require reference electrodes or involve complex fabrication processes. A recently proposed bulk junctionless BioFET (Bulk JL-BioFET) features a simple fabrication process to address these issues. This structure utilizes a depletion region formed by a p-n junction, as the active layer is directly in contact with a substrate of the opposite type. As a result, the device can operate effectively with only two terminals—drain and source—without the need for a reference electrode. In this study, we propose a novel Bulk JL-BioFET, incorporating a doped field stop layer and an asymmetric source/drain structure, and verify its performance through simulations. The doped field stop layer blocks the electric field expansion, enhancing channel modulation, while the asymmetric source/drain structure promotes electron injection, reducing the on-off swing voltage and turn-on voltage. This improves the electrical performance, enabling lower power consumption and higher sensitivity. Simulation results show that the combination of these two novel features results in a sensitivity increase of approximately 30-fold. Moreover, high sensitivity was observed below the turn-on voltage region for all the structures when analyzing the sensitivity with overdrive voltage, identifying the optimal operating conditions. This study suggests that the combination of the doped field stop layer and asymmetric source/drain structure is an effective design strategy to maximize the sensing performance of BioFETs while minimizing power consumption.
Full article
(This article belongs to the Special Issue Transistor-Based Biosensors and Their Applications)
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Open AccessArticle
Pathogen-on-a-Chip: Impedance-Based Detection of Biofilm Formation of Staphylococcus aureus and Staphylococcus epidermidis
by
Bengisu Yöney, Radka Obořilová, Karel Lacina, Zdeněk Farka and Petr Skládal
Biosensors 2025, 15(9), 596; https://doi.org/10.3390/bios15090596 - 10 Sep 2025
Abstract
Bacterial biofilms are complex microbial communities that contribute to the pathogenesis of chronic infections. Therefore, it is crucial to detect biofilm-associated infections in early stages as their delayed treatment becomes more complicated. Herein, we describe a label-free electrochemical impedance spectroscopy (EIS) method for
[...] Read more.
Bacterial biofilms are complex microbial communities that contribute to the pathogenesis of chronic infections. Therefore, it is crucial to detect biofilm-associated infections in early stages as their delayed treatment becomes more complicated. Herein, we describe a label-free electrochemical impedance spectroscopy (EIS) method for detecting biofilm formation by Staphylococcus aureus and Staphylococcus epidermidis. Printed circuit board-based biamperometric gold electrodes were modified with poly-L-lysine to enhance bacterial attachment to the sensor surface. Formation and inhibition of biofilms were evaluated based on changes in charge transfer resistance (Rct). The control Rct value increased by ~90 kΩ for S. epidermidis biofilm and by ~60 kΩ for S. aureus biofilms. Antibiotic-treated samples exhibited similar values to those using the control. In addition, biofilm formation was evaluated through optical microscopy using safranin staining, and the micrographs suggest significant biomass on the electrodes, whereas the control appeared clear. Atomic force microscopy was used to visualize the biofilm on the electrode surface, obtain cross-sectional profiles, and evaluate its roughness. The roughness parameters indicate that S. aureus forms a rougher biofilm than S. epidermidis, while S. epidermidis forms a more compact biofilm. These findings suggest that the optimized EIS-based method effectively monitors changes related to biofilms and serves as a promising tool for evaluation of new anti-biofilm agents, such as antibiotics, phages or antibodies.
Full article
(This article belongs to the Special Issue Electrochemical (Bio)Sensors as Promising Analytical Tools in the Analysis of Soils, Plants and Environmental Monitoring)
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Open AccessReview
Advancements and Applications of Split Technology in CRISPR/Cas12a: Transforming Molecular Diagnostics and Biosensing
by
Saikarthik Jayakumar, Srinivasan Vengadassalapathy, Santhosh Venkadassalapathy, Sheela Durairajan, Radha Vijayaraj and Lakshmanan Govindan
Biosensors 2025, 15(9), 595; https://doi.org/10.3390/bios15090595 - 10 Sep 2025
Abstract
The rapid evolution of CRISPR technology has revolutionized molecular biology, and among the various systems, CRISPR/Cas12a stands out for its high specificity and efficient collateral cleavage activity. This review article focuses on the recent advancements and applications of split technology within the CRISPR/Cas12a
[...] Read more.
The rapid evolution of CRISPR technology has revolutionized molecular biology, and among the various systems, CRISPR/Cas12a stands out for its high specificity and efficient collateral cleavage activity. This review article focuses on the recent advancements and applications of split technology within the CRISPR/Cas12a framework, highlighting its transformative role in molecular diagnostics and biosensing. Split technology innovatively divides functional nucleic acid components into modular segments that are activated by specific targets, significantly enhancing the specificity and sensitivity of biosensors. This design addresses the inherent limitations of traditional sensor systems, enabling the direct detection of ultrashort nucleic acids and improved discrimination of single-nucleotide variants, thereby facilitating the simultaneous detection of multiple biomolecules. The versatility of split-enabled biosensors extends beyond genetic testing, making them valuable tools in diagnostics, therapeutics, and environmental science. Despite challenges such as crRNA degradation and reassembly kinetics, ongoing research and engineering solutions continue to enhance the stability and performance of these systems. This review synthesizes the foundational principles, recent advancements, and potential applications of split technology while also identifying challenges and opportunities for future exploration. Ultimately, our insights provide a comprehensive resource to leverage the full potential of CRISPR/Cas12a-based split technology in advancing biosensing methodologies and clinical applications.
Full article
(This article belongs to the Section Biosensor and Bioelectronic Devices)
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Open AccessArticle
Whole-Cell Bioreporter-Based Assay for Detecting Fungal-Derived β-Lactamase Inhibitors
by
Raz Benou, Robert S. Marks, Alex Sivan, Esti Kramarsky-Winter, Karina Golberg and Ariel Kushmaro
Biosensors 2025, 15(9), 594; https://doi.org/10.3390/bios15090594 - 9 Sep 2025
Abstract
β-lactams are an important family of antibiotics that are prone to undergo resistance inhibition though the production of β-lactamases by some microorganisms. To combat this resistance and preserve the efficacy of β-lactam antibiotics, we developed a strategy for the discovery of such β-lactamase
[...] Read more.
β-lactams are an important family of antibiotics that are prone to undergo resistance inhibition though the production of β-lactamases by some microorganisms. To combat this resistance and preserve the efficacy of β-lactam antibiotics, we developed a strategy for the discovery of such β-lactamase inhibitors. When combined with β-lactams, these inhibitors allow the antibiotics to be effective and prevent resistance. To date, the development of such combinatory drugs is limited due to the complexity of screening for new β-lactamase inhibitors. Therefore, to facilitate this development, it was essential to find sensitive assays to effectively screen for lactamase inhibitory compounds. To this end, a novel bioassay utilizing bioluminescent indicator bacteria as bioreporters was developed. The assay was first optimized using commercial antibiotics together with known β-lactamase inhibitors. Using this bioassay, we then screened for novel natural β-lactamase inhibitors derived from coral-associated fungi. We showed that the fungus Penicillium spinulosum, originating from the coral Pocillopora sp. from the Gulf of Aqaba Eilat, produced compounds with anti-β-lactamase activity. We further demonstrated that the bioreporter bacteria used here responded to the combined antibiotics and β-lactamase inhibitors in a concentration-dependent manner, indicating their usefulness for β-lactamase-inhibiting compound discovery. Future structural identification will promote the validation of this assay’s usefulness.
Full article
(This article belongs to the Section Biosensor and Bioelectronic Devices)
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Open AccessReview
Role of Polymeric Stabilizing Agents as a Molecular Spacer in Gold Nanoparticle-Mediated FRET-Based Biosensing
by
Atul Kumar Tiwari and Roger J. Narayan
Biosensors 2025, 15(9), 593; https://doi.org/10.3390/bios15090593 - 9 Sep 2025
Abstract
In FRET-based sensing, the interaction between the analytes and metal nanoparticles is significantly regulated by the physicochemical characteristics of the nanoparticles, such as their shape, size, zeta potential, surface-linked ligands, doping, pH of the medium, particle surface roughness, and lattice structure (atomic arrangements).
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In FRET-based sensing, the interaction between the analytes and metal nanoparticles is significantly regulated by the physicochemical characteristics of the nanoparticles, such as their shape, size, zeta potential, surface-linked ligands, doping, pH of the medium, particle surface roughness, and lattice structure (atomic arrangements). During the synthesis process, to avert the aggregation of gold nanoparticles (AuNPs), synthetic polymers (including polyethylene glycol, polyethyleneimine, and poly-N-vinylpyrrolidone) and natural polymers (such as chitosan, starch, gellan, welan, and κ-carrageenan) are frequently employed for stabilization. This stabilization is accomplished through mechanisms such as steric repulsion and electrostatic stabilization, which form a protective layer around AuNPs. These stabilizing polymers act as molecular spacers in nanoparticle-based FRET sensing, enabling the precise regulation of the molecular distance between the acceptor and donor fluorophore molecules. This regulation enhances the efficiency and sensitivity of FRET assays. By modifying the length and flexibility of the spacer polymer, researchers can adjust the spacing between fluorophores, ensuring effective energy transfer and the accurate detection of target molecules. However, there is a limited understanding of the role and broad application of these molecular spacers in nanoparticle-mediated FRET-based sensing of various analytes. Consequently, this review explores different fundamental aspects of FRET, polymeric stabilization of gold nanoparticles, and various polymeric spacers in FRET-based sensing, along with the recent advancements and challenges associated with this approach.
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(This article belongs to the Special Issue Advances in Portable and Wearable Sensing Systems for Biochemical Monitoring)
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Open AccessArticle
Development of a Multiplex Lateral Flow Immunoassay for the Detection of Antibiotics in Milk Utilizing Lyophilized Gold Nanoparticle Conjugates
by
Ivan V. Maksin, Azhar Kuandykova, Darya I. Polyakova, Viktoriia A. Kesareva, Timofei A. Luzyanin, Vladislav S. Ivanov, Evgeniia I. Simonova, German A. Khunteev and Yuliya G. Kirillova
Biosensors 2025, 15(9), 592; https://doi.org/10.3390/bios15090592 - 9 Sep 2025
Abstract
Lateral flow immunoassays (LFAs) are widely recognized as a powerful and versatile analytical platform. Nevertheless, the development of multiplex formats remains a distinct challenge. The aim of this study was to develop a multiplex LFA using gold nanoparticles (GNPs) as a label, selected
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Lateral flow immunoassays (LFAs) are widely recognized as a powerful and versatile analytical platform. Nevertheless, the development of multiplex formats remains a distinct challenge. The aim of this study was to develop a multiplex LFA using gold nanoparticles (GNPs) as a label, selected for their ease of synthesis and functionalization with biomolecules. We provide practical recommendations regarding protein–hapten synthesis, membrane selection, application buffer composition, and methods to improve the long-term stability of the freeze-dried gold conjugate. The developed assay shows good tolerance to high-fat milk, stability at elevated temperatures, and promising sensitivity, with visual detection limits of 4–100 ng/mL for -lactams, 1–10 ng/mL for tetracyclines, 50 ng/mL for streptomycin, and 0.3 ng/mL for chloramphenicol.
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(This article belongs to the Special Issue Immunosensors: Design and Applications)
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Open AccessReview
Review of Pulsation Signal Detection and Applications in Dynamic Photoacoustic Imaging
by
Wenhan Zheng, Chuqin Huang and Jun Xia
Biosensors 2025, 15(9), 591; https://doi.org/10.3390/bios15090591 - 8 Sep 2025
Abstract
Pulsatile signal detection plays an important role in monitoring various physiological parameters, primarily heart rate and blood oxygen saturation. Their applications range from clinical settings to personal health and wellness monitoring. PPG (photoplethysmography) can provide non-invasive optical measurements to detect blood volume changes
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Pulsatile signal detection plays an important role in monitoring various physiological parameters, primarily heart rate and blood oxygen saturation. Their applications range from clinical settings to personal health and wellness monitoring. PPG (photoplethysmography) can provide non-invasive optical measurements to detect blood volume changes in peripheral tissues. Yet, it suffers from low spatial resolution to precisely detect the pulsatile signal originating over 2 mm in human tissue. Ultrasound (US) provides a deep detectable range compared to the pure optical method. However, its low contrast to red blood cells and cluster artifacts makes it only detect the indirect pulsation from the surrounding tissue of blood vessels. Recent advances in PA imaging show its capability to precisely measure pulsatile signals originating from blood vessels in deep regions (over 10 mm) and its potential to accurately record blood oxygen saturation with high spatial and temporal resolution. This review article summarizes studies on photoacoustic (PA) pulsatile signal monitoring, highlights the technical advances, and compares it against optical and ultrasonic approaches.
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(This article belongs to the Special Issue Advanced Optical Methods for Biosensing)
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Open AccessSystematic Review
Natural Iron Oxide Nanoparticles Produced by Aquatic Magnetotactic Bacteria as Ideal Nanozymes for Nano-Guided Biosensing Platforms—A Systematic Review
by
Natalia Lorela Paul, Catalin Ovidiu Popa and Rodica Elena Ionescu
Biosensors 2025, 15(9), 590; https://doi.org/10.3390/bios15090590 - 8 Sep 2025
Abstract
In response to the ongoing challenges associated with natural enzymes, their high production costs, low stability and limited functionality; nanozymes have rapidly emerged as versatile alternative. Such nanocatalysts, based on nanomaterials and nanostructures, offer remarkable tunability of physicochemical properties and excellent durability, and
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In response to the ongoing challenges associated with natural enzymes, their high production costs, low stability and limited functionality; nanozymes have rapidly emerged as versatile alternative. Such nanocatalysts, based on nanomaterials and nanostructures, offer remarkable tunability of physicochemical properties and excellent durability, and adapt themselves effectively to the requirements of modern biotechnological applications. This review article aims to provide a comprehensive overview of recent advances in the use of naturally occurring iron oxide nanoparticles, produced by magnetotactic bacteria, and to highlight their emerging role as key elements in the development of the new generation of nano-guided biosensors. It provides a comprehensive and systematic analysis of publications in the Web of Science database between 2022 and August 2025, conducted in accordance with PRISMA guidelines. The aim was to assess the current state of the art and identify knowledge gaps in the exploration and application of magnetotactic bacteria as natural and sustainable sources in the design of next-generation biosensors. The natural nanoparticles, formed through biological processes, represent a unique and sustainable alternative to synthetic nanoparticles, offering naturally mimetic enzymatic activity, high biocompatibility, and exceptional stability. This approach opens up revolutionary perspectives in the field of biosensors, proposing a new class of functional materials, iron nanoparticles of biological origin, capable of fundamentally changing the performance, sustainability and reliability of future nanoenzymatic sensing platforms.
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(This article belongs to the Special Issue Cutting-Edge Nanozyme Biosensing Strategies for Biomedical and Environmental Applications)
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Open AccessReview
POC Sensor Systems and Artificial Intelligence—Where We Are Now and Where We Are Going?
by
Prashanthi Kovur, Krishna M. Kovur, Dorsa Yahya Rayat and David S. Wishart
Biosensors 2025, 15(9), 589; https://doi.org/10.3390/bios15090589 - 8 Sep 2025
Abstract
Integration of machine learning (ML) and artificial intelligence (AI) into point-of-care (POC) sensor systems represents a transformative advancement in healthcare. This integration enables sophisticated data analysis and real-time decision-making in emergency and intensive care settings. AI and ML algorithms can process complex biomedical
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Integration of machine learning (ML) and artificial intelligence (AI) into point-of-care (POC) sensor systems represents a transformative advancement in healthcare. This integration enables sophisticated data analysis and real-time decision-making in emergency and intensive care settings. AI and ML algorithms can process complex biomedical data, improve diagnostic accuracy, and enable early disease detection for better patient outcomes. Predictive analytics in POC devices supports proactive healthcare by analyzing data to forecast health issues and facilitating early intervention and personalized treatment. This review covers the key areas of ML and AI integration in POC devices, including data analysis, pattern recognition, real-time decision support, predictive analytics, personalization, automation, and workflow optimization. Examples of current POC devices that use ML and AI include AI-powered blood glucose monitors, portable imaging devices, wearable cardiac monitors, AI-enhanced infectious disease detection, and smart wound care sensors are also discussed. The review further explores new directions for POC sensors and ML integration, including mental health monitoring, nutritional monitoring, metabolic health tracking, and decentralized clinical trials (DCTs). We also examined the impact of integrating ML and AI into POC devices on healthcare accessibility, efficiency, and patient outcomes.
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(This article belongs to the Special Issue Trends in Development of Biosensors for Disease Diagnosis, Treatment and Management—2nd Edition)
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