Biosensors doi: 10.3390/bios14040161
Authors: Monkrathok Janphuang Suphachiaraphan Kampaengsri Kamkaew Chansaenpak Lisnund Blay Pinyou
We designed and optimized a glucose biosensor system based on a screen-printed electrode modified with the NAD-GDH enzyme. To enhance the electroactive surface area and improve the electron transfer efficiency, we introduced graphene oxide (GO) and ferrocene-modified linear poly(ethylenimine) (LPEI-Fc) onto the biosensor surface. This strategic modification exploits the electrostatic interaction between graphene oxide, which possesses a negative charge, and LPEI-Fc, which is positively charged. This interaction results in increased catalytic current during glucose oxidation and helps improve the overall glucose detection sensitivity by amperometry. We integrated the developed glucose sensor into a flow injection (FI) system. This integration facilitates a swift and reproducible detection of glucose, and it also mitigates the risk of contamination during the analyses. The incorporation of an FI system improves the efficiency of the biosensor, ensuring precise and reliable results in a short time. The proposed sensor was operated at a constant applied potential of 0.35 V. After optimizing the system, a linear calibration curve was obtained for the concentration range of 1.0–40 mM (R2 = 0.986). The FI system was successfully applied to determine the glucose content of a commercial sports drink.
]]>Biosensors doi: 10.3390/bios14040160
Authors: Ran Bi Jianxiong Yang Chengqi Huang Xiaoyu Zhang Ran Liao Hui Ma
Harmful algal blooms (HABs) pose a global threat to the biodiversity and stability of local aquatic ecosystems. Rapid and accurate classification of microalgae and cyanobacteria in water is increasingly desired for monitoring complex water environments. In this paper, we propose a pulse feature-enhanced classification (PFEC) method as a potential solution. Equipped with a rapid measurement prototype that simultaneously detects polarized light scattering and fluorescence signals of individual particles, PFEC allows for the extraction of 38 pulse features to improve the classification accuracy of microalgae, cyanobacteria, and other suspended particulate matter (SPM) to 89.03%. Compared with microscopic observation, PFEC reveals three phyla proportions in aquaculture samples with an average error of less than 14%. In this paper, PFEC is found to be more accurate than the pulse-average classification method, which is interpreted as pulse features carrying more detailed information about particles. The high consistency of the dominant and common species between PFEC and microscopy in all field samples also demonstrates the flexibility and robustness of the former. Moreover, the high Pearson correlation coefficient accounting for 0.958 between the cyanobacterial proportion obtained by PFEC and the cyanobacterial density given by microscopy implies that PFEC serves as a promising early warning tool for cyanobacterial blooms. The results of this work suggest that PFEC holds great potential for the rapid and accurate classification of microalgae and cyanobacteria in aquatic environment monitoring.
]]>Biosensors doi: 10.3390/bios14040159
Authors: Diogo Costa Patrícia Pereira-Silva Paulo Sousa Vânia Pinto Joel Borges Filipe Vaz Graça Minas Paula Sampaio
This work reports on the surface functionalization of a nanomaterial supporting localized surface plasmon resonances (LSPRs) with (synthetic) thiolated oligonucleotide-based biorecognition elements, envisaging the development of selective LSPR-based DNA biosensors. The LSPR thin-film transducers are composed of noble metal nanoparticles (NPs) embedded in a TiO2 dielectric matrix, produced cost-effectively and sustainably by magnetron sputtering. The study focused on the immobilization kinetics of thiolated oligonucleotide probes as biorecognition elements, followed by the evaluation of hybridization events with the target probe. The interaction between the thiolated oligonucleotide probe and the transducer’s surface was assessed by monitoring the LSPR signal with successive additions of probe solution through a microfluidic device. The device was specifically designed and fabricated for this work and adapted to a high-resolution LSPR spectroscopy system with portable characteristics. Benefiting from the synergetic characteristics of Ag and Au in the form of bimetallic nanoparticles, the Au-Ag/TiO2 thin film proved to be more sensitive to thiolated oligonucleotide binding events. Despite the successful surface functionalization with the biorecognition element, the detection of complementary oligonucleotides revealed electrostatic repulsion and steric hindrance, which hindered hybridization with the target oligonucleotide. This study points to an effect that is still poorly described in the literature and affects the design of LSPR biosensors based on nanoplasmonic thin films.
]]>Biosensors doi: 10.3390/bios14040158
Authors: Zhishan Yuan Yanbang Lin Jieming Hu Chengyong Wang
Solid-state nanopores have become a prominent tool in the field of single-molecule detection. Conventional solid-state nanopores are thick, which affects the spatial resolution of the detection results. Graphene is the thinnest 2D material and has the highest spatial detection resolution. In this study, a graphene membrane chip was fabricated by combining a MEMS process with a 2D material wet transfer process. Raman spectroscopy was used to assess the quality of graphene after the transfer. The mechanism behind the influence of the processing dose and residence time of the helium ion beam on the processed pore size was investigated. Subsequently, graphene nanopores with diameters less than 10 nm were fabricated via helium ion microscopy. DNA was detected using a 5.8 nm graphene nanopore chip, and the appearance of double-peak signals on the surface of 20 mer DNA was successfully detected. These results serve as a valuable reference for nanopore fabrication using 2D material for DNA analysis.
]]>Biosensors doi: 10.3390/bios14040157
Authors: Amy Makler-Disatham Massimo Caputi Waseem Asghar
Since its first appearance in 1981, HIV-1 has remained a global concern. Current methods for diagnosing HIV-1, while effective, are mostly specific to a given subtype of HIV-1 and often require expensive equipment and highly trained individuals to collect and process the sample. It is necessary to develop a sensitive diagnostic method that can be administered with minimal equipment to provide better care in low-resource settings. Loop-mediated isothermal amplification is a rapid and sensitive method for detecting the presence of specific nucleic acid sequences. Herein we report the development and comparison of two different HIV LAMP assays, integrase and VPR, as well as the comparison between TRIZol and magnetic beads RNA extraction methods for each assay. Our analysis shows that the integrase assay was able to detect the virus from multiple subtypes in under 30 min with a variable limit of detection (LOD) that was dependent on the HIV-1 subtype.
]]>Biosensors doi: 10.3390/bios14040156
Authors: Jessica Petiti Laura Revel Carla Divieto
The resazurin assay, also known as the Alamar Blue assay, stands as a cornerstone technique in cell biology, microbiology, and drug development. It assesses the viability of cells through the conversion of resazurin into highly fluorescent resorufin. The resulting fluorescence intensity provides a reliable estimate of viable cell numbers. Cytotoxicity assays, such as the resazurin-based method, play a crucial role in the screening of potential drug candidates and in the assessment of pharmaceutical and chemical toxicity. In recent years, inconsistencies have arisen in pharmacogenomic studies, often due to poorly optimized laboratory protocols. These inconsistencies hinder progress in understanding how substances affect cell health, leading to unreliable findings. Thus, the need for standardized and rigorously optimized protocols is evident to ensure consistent and accurate results in cytotoxicity studies. This manuscript describes a standardized procedure for optimizing resazurin-based viability assays to improve the reliability of cytotoxicity data. This optimization approach focuses on critical experimental parameters and data quality, aiming to achieve a level of measurement imprecision of less than 20%. In conclusion, to address the critical issues of reproducibility and reliability, protocol standardization, such as the one described in this manuscript, can greatly enhance the credibility of cytotoxicity studies, ultimately advancing drug safety assessments.
]]>Biosensors doi: 10.3390/bios14040155
Authors: Hideto Kumagai Hiroyuki Furusawa
Nucleic acid amplification reactions such as polymerase chain reaction (PCR), which uses a DNA polymerase to amplify individual double-stranded DNA fragments, are a useful technique for visualizing the presence of specific genomes. Although the fluorescent labeling method is mainly used with DNA amplification, other detection methods should be considered for further improvements, such as miniaturization and cost reduction, of reaction-monitoring devices. In this study, the quartz-crystal microbalance (QCM) method, which can measure nanogram-order masses, was applied for the real-time detection of DNA fragments in a solution with nucleic acids. This was combined with an isothermal nucleic acid amplification reaction based on the recombinase polymerase amplification (RPA) method, which allowed DNA amplification at a constant temperature. When the DNA amplification reaction was initiated on a QCM sensor plate with an immobilized primer DNA strand, a significant increase in mass was observed compared to when the primer DNA was not immobilized. QCM was shown to be sufficiently sensitive for the in situ detection of amplified DNA fragments. Combining a portable QCM device and RPA offers a sensitive point-of-care method for detecting nucleic acids.
]]>Biosensors doi: 10.3390/bios14040154
Authors: Filippo Storti Silvio Bonfadini Gaia Bondelli Vito Vurro Guglielmo Lanzani Luigino Criante
Measuring the transit time of a cell forced through a bottleneck is one of the most widely used techniques for the study of cell deformability in flow. It in turn provides an accessible and rapid way of obtaining crucial information regarding cell physiology. Many techniques are currently being investigated to reliably retrieve this time, but their translation to diagnostic-oriented devices is often hampered by their complexity, lack of robustness, and the bulky external equipment required. Herein, we demonstrate the benefits of coupling microfluidics with an optical method, like photocells, to measure the transit time. We exploit the femtosecond laser irradiation followed by chemical etching (FLICE) fabrication technique to build a monolithic 3D device capable of detecting cells flowing through a 3D non-deformable constriction which is fully buried in a fused silica substrate. We validated our chip by measuring the transit times of pristine breast cancer cells (MCF-7) and MCF-7 cells treated with Latrunculin A, a drug typically used to increase their deformability. A difference in transit times can be assessed without the need for complex external instrumentation and/or demanding computational efforts. The high throughput (4000–10,000 cells/min), ease of use, and clogging-free operation of our device bring this approach much closer to real scenarios.
]]>Biosensors doi: 10.3390/bios14030153
Authors: Kiyoto Sanjo Kazuki Hebiguchi Cheng Tang Essam A. Rashed Sachiko Kodera Hiroyoshi Togo Akimasa Hirata
An electrocardiogram (ECG) is used to observe the electrical activity of the heart via electrodes on the body surface. Recently, an ECG with fewer electrodes, such as a bipolar ECG in which two electrodes are attached to the chest, has been employed as wearable devices. However, the effect of different geometrical factors and electrode-pair locations on the amplitude and waveform of ECG signals remains unclear. In this study, we computationally evaluated the effects of body morphology, heart size and orientation, and electrode misalignment on ECG signals for 48 scenarios using 35 bipolar electrode pairs (1680 waveforms) with a dynamic time warping (DTW) algorithm. It was observed that the physique of the human body model predominantly affected the amplitude and waveform of the ECG signals. A multivariate analysis indicated that the heart–electrode distance and the solid angle of the heart from the electrode characterized the amplitude and waveform of the ECG signals, respectively. Furthermore, the electrode locations for less individual variability and less waveform distortion were close to the location of electrodes V2 and V3 in the standard 12-lead. These findings will facilitate the placement of ECG electrodes and interpretation of the measured ECG signals for wearable devices.
]]>Biosensors doi: 10.3390/bios14030152
Authors: Juhee Lim Won Han Le Tran Huy Thang Yong Wook Lee Joong Ho Shin
Accurate sample heating is vital for nucleic acid extraction and amplification, requiring a sophisticated thermal cycling process in nucleic acid detection. Traditional molecular detection systems with heating capability are bulky, expensive, and primarily designed for lab settings. Consequently, their use is limited where lab systems are unavailable. This study introduces a technique for performing the heating process required in molecular diagnostics applicable for point-of-care testing (POCT), by presenting a method for crafting customized heaters using freely patterned nichrome (NiCr) wire. This technique, fabricating heaters by arranging protrusions on a carbon black-polydimethylsiloxane (PDMS) cast and patterning NiCr wire, utilizes cost-effective materials and is not constrained by shape, thereby enabling customized fabrication in both two-dimensional (2D) and three-dimensional (3D). To illustrate its versatility and practicality, a 2D heater with three temperature zones was developed for a portable device capable of automatic thermocycling for polymerase chain reaction (PCR) to detect Escherichia coli (E. coli) O157:H7 pathogen DNA. Furthermore, the detection of the same pathogen was demonstrated using a customized 3D heater surrounding a microtube for loop-mediated isothermal amplification (LAMP). Successful DNA amplification using the proposed heater suggests that the heating technique introduced in this study can be effectively applied to POCT.
]]>Biosensors doi: 10.3390/bios14030151
Authors: Aya M. Mostafa Stephen J. Barton Stephen P. Wren James Barker
Two fluorescent molecularly imprinted polymers (MIPs) were developed for pepsin enzyme utilising fluorescein and rhodamine b. The main difference between both dyes is the presence of two (diethylamino) groups in the structure of rhodamine b. Consequently, we wanted to investigate the effect of these functional groups on the selectivity and sensitivity of the resulting MIPs. Therefore, two silica-based MIPs for pepsin enzyme were developed using 3-aminopropyltriethoxysilane as a functional monomer and tetraethyl orthosilicate as a crosslinker to achieve a one-pot synthesis. Results of our study revealed that rhodamine b dyed MIPs (RMIPs) showed stronger binding, indicated by a higher binding capacity value of 256 mg g−1 compared to 217 mg g−1 for fluorescein dyed MIPs (FMIPs). Moreover, RMIPs showed superior sensitivity in the detection and quantitation of pepsin with a linear range from 0.28 to 42.85 µmol L−1 and a limit of detection (LOD) as low as 0.11 µmol L−1. In contrast, FMIPs covered a narrower range from 0.71 to 35.71 µmol L−1, and the LOD value reached 0.34 µmol L−1, which is three times less sensitive than RMIPs. Finally, the developed FMIPs and RMIPs were applied to a separation-free quantification system for pepsin in saliva samples without interference from any cross-reactors.
]]>Biosensors doi: 10.3390/bios14030150
Authors: Sankar Prasad Bag Suyoung Lee Jaeyoon Song Jinsink Kim
Hydrogel-gated synaptic transistors offer unique advantages, including biocompatibility, tunable electrical properties, being biodegradable, and having an ability to mimic biological synaptic plasticity. For processing massive data with ultralow power consumption due to high parallelism and human brain-like processing abilities, synaptic transistors have been widely considered for replacing von Neumann architecture-based traditional computers due to the parting of memory and control units. The crucial components mimic the complex biological signal, synaptic, and sensing systems. Hydrogel, as a gate dielectric, is the key factor for ionotropic devices owing to the excellent stability, ultra-high linearity, and extremely low operating voltage of the biodegradable and biocompatible polymers. Moreover, hydrogel exhibits ionotronic functions through a hybrid circuit of mobile ions and mobile electrons that can easily interface between machines and humans. To determine the high-efficiency neuromorphic chips, the development of synaptic devices based on organic field effect transistors (OFETs) with ultra-low power dissipation and very large-scale integration, including bio-friendly devices, is needed. This review highlights the latest advancements in neuromorphic computing by exploring synaptic transistor developments. Here, we focus on hydrogel-based ionic-gated three-terminal (3T) synaptic devices, their essential components, and their working principle, and summarize the essential neurodegenerative applications published recently. In addition, because hydrogel-gated FETs are the crucial members of neuromorphic devices in terms of cutting-edge synaptic progress and performances, the review will also summarize the biodegradable and biocompatible polymers with which such devices can be implemented. It is expected that neuromorphic devices might provide potential solutions for the future generation of interactive sensation, memory, and computation to facilitate the development of multimodal, large-scale, ultralow-power intelligent systems.
]]>Biosensors doi: 10.3390/bios14030149
Authors: Andrea Martínez-Lozano Roberto Gutierrez Carlos G. Juan Carolina Blanco-Angulo Héctor García-Martínez Germán Torregrosa José María Sabater-Navarro Ernesto Ávila-Navarro
A proof-of-concept of a microwave imaging system for the fast detection of abdominal aortic aneurysms is shown. This experimental technology seeks to overcome the factors hampering the fast screening for these aneurysms with the usual equipment, such as high cost, long-time operation or hazardous exposure to chemical substances. The hardware system is composed of 16 twin antennas mastered by a microcontroller through a switching network, which connects the antennas to the measurement instrument for sequential measurement. The software system is run by a computer, mastering the whole system, automatizing the measurement process and running the signal processing and medical image generation algorithms. Two image generation algorithms are tested: Delay-and-Sum (DAS) and Improved Delay-and-Sum (IDAS). Own-modified versions of these algorithms adapted to the requirements of our system are proposed. The system is carefully calibrated and fine-tuned with known objects placed at known distances. An experimental proof-of-concept is shown with a human torso phantom, including an aorta phantom and an aneurysm phantom placed in different positions. The results show good imaging capabilities with the potential for detecting and locating possible abdominal aortic aneurysms and reporting acceptable errors.
]]>Biosensors doi: 10.3390/bios14030148
Authors: Jose-Luis Lafuente Samuel González Clara Aibar Desirée Rivera Eva Avilés Juan-Jose Beunza
Lactate, once merely regarded as an indicator of tissue hypoxia and muscular fatigue, has now gained prominence as a pivotal biomarker across various medical disciplines. Recent research has unveiled its critical role as a high-value prognostic marker in critical care medicine. The current practice of lactate detection involves periodic blood sampling. This approach is invasive and confined to measurements at six-hour intervals, leading to resource expenditure, time consumption, and patient discomfort. This review addresses non-invasive sensors that enable continuous monitoring of lactate in critical care patients. After the introduction, it discusses the iontophoresis system, followed by a description of the structural materials that are universally employed to create an interface between the integumentary system and the sensor. Subsequently, each method is detailed according to its physical principle, outlining its advantages, limitations, and pertinent aspects. The study concludes with a discussion and conclusions, aiming at the design of an intelligent sensor (Internet of Medical Things or IoMT) to facilitate continuous lactate monitoring and enhance the clinical decision-making support system in critical care medicine.
]]>Biosensors doi: 10.3390/bios14030147
Authors: Jiechang Guo Aygun Teymur Chenling Tang Ramesh Saxena Tianfu Wu
To improve the efficiency and patient coverage of the current healthcare system, user-friendly novel homecare devices are urgently needed. In this work, we developed a smartphone-based analyzing and reporting system (SBARS) for biomarker detection in lupus nephritis (LN). This system offers a cost-effective alternative to traditional, expensive large equipment in signal detection and quantification. This innovative approach involves using a portable and affordable microscopic reader to capture biomarker signals. Through smartphone-based image processing techniques, the intensity of each biomarker signal is analyzed. This system exhibited comparable performance to a commercial Genepix scanner in the detection of two potential novel biomarkers of LN, VISG4 and TNFRSF1b. Importantly, this smartphone-based analyzing and reporting system allows for discriminating LN patients with active renal disease from healthy controls with the area-under-the-curve (AUC) value = 0.9 for TNFRSF1b and 1.0 for VSIG4, respectively, indicating high predictive accuracy.
]]>Biosensors doi: 10.3390/bios14030146
Authors: Ki Sung Park Tae-In Park Jae Eon Lee Seo-Yeong Hwang Anna Choi Seung Pil Pack
The global challenges posed by the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) pandemic have underscored the critical importance of innovative and efficient control systems for addressing future pandemics. The most effective way to control the pandemic is to rapidly suppress the spread of the virus through early detection using a rapid, accurate, and easy-to-use diagnostic platform. In biosensors that use bioprobes, the binding affinity of molecular recognition elements (MREs) is the primary factor determining the dynamic range of the sensing platform. Furthermore, the sensitivity relies mainly on bioprobe quality with sufficient functionality. This comprehensive review investigates aptamers and nanobodies recently developed as advanced MREs for SARS-CoV-2 diagnostic and therapeutic applications. These bioprobes might be integrated into organic bioelectronic materials and devices, with promising enhanced sensitivity and specificity. This review offers valuable insights into advancing biosensing technologies for infectious disease diagnosis and treatment using aptamers and nanobodies as new bioprobes.
]]>Biosensors doi: 10.3390/bios14030145
Authors: Xin Li Zhisheng Dang Wenqiang Tang Haoji Zhang Jianwei Shao Rui Jiang Xu Zhang Fuqiang Huang
The rapid and accurate identification of parasites is crucial for prompt therapeutic intervention in parasitosis and effective epidemiological surveillance. For accurate and effective clinical diagnosis, it is imperative to develop a nucleic-acid-based diagnostic tool that combines the sensitivity and specificity of nucleic acid amplification tests (NAATs) with the speed, cost-effectiveness, and convenience of isothermal amplification methods. A new nucleic acid detection method, utilizing the clustered regularly interspaced short palindromic repeats (CRISPR)-associated (Cas) nuclease, holds promise in point-of-care testing (POCT). CRISPR/Cas12a is presently employed for the detection of Plasmodium falciparum, Toxoplasma gondii, Schistosoma haematobium, and other parasites in blood, urine, or feces. Compared to traditional assays, the CRISPR assay has demonstrated notable advantages, including comparable sensitivity and specificity, simple observation of reaction results, easy and stable transportation conditions, and low equipment dependence. However, a common issue arises as both amplification and cis-cleavage compete in one-pot assays, leading to an extended reaction time. The use of suboptimal crRNA, light-activated crRNA, and spatial separation can potentially weaken or entirely eliminate the competition between amplification and cis-cleavage. This could lead to enhanced sensitivity and reduced reaction times in one-pot assays. Nevertheless, higher costs and complex pre-test genome extraction have hindered the popularization of CRISPR/Cas12a in POCT.
]]>Biosensors doi: 10.3390/bios14030142
Authors: Nathan Vanalken Katrijn Boon Martyna Szpakowska Andy Chevigné Dominique Schols Tom Van Loy
The human CC chemokine receptor 7 (CCR7) is activated by two natural ligands, CC chemokine ligand 19 (CCL19) and 21 (CCL21). The CCL19-CCL21-CCR7 axis has been extensively studied in vitro, but there is still debate over whether CCL21 is an overall weaker agonist or if the axis displays biased signalling. In this study, we performed a systematic analysis at the transducer level using NanoBRET-based methodologies in three commonly used cellular backgrounds to evaluate pathway and ligand preferences, as well as ligand bias and the influence of the cellular system thereon. We found that both CCL19 and CCL21 activated all cognate G proteins and some non-cognate couplings in a cell-type-dependent manner. Both ligands recruited β-arrestin1 and 2, but the potency was strongly dependent on the cellular system. Overall, CCL19 and CCL21 showed largely conserved pathway preferences, but small differences were detected. However, these differences only consolidated in a weak ligand bias. Together, these data suggest that CCL19 and CCL21 share mostly overlapping, weakly biased, transducer profiles, which can be influenced by the cellular context.
]]>Biosensors doi: 10.3390/bios14030143
Authors: Cristina Purcarea Robert Ruginescu Roberta Maria Banciu Alina Vasilescu
Extremozymes combine high specificity and sensitivity with the ability to withstand extreme operational conditions. This work presents an overview of extremozymes that show potential for environmental monitoring devices and outlines the latest advances in biosensors utilizing these unique molecules. The characteristics of various extremozymes described so far are presented, underlining their stability and operational conditions that make them attractive for biosensing. The biosensor design is discussed based on the detection of photosynthesis-inhibiting herbicides as a case study. Several biosensors for the detection of pesticides, heavy metals, and phenols are presented in more detail to highlight interesting substrate specificity, applications or immobilization methods. Compared to mesophilic enzymes, the integration of extremozymes in biosensors faces additional challenges related to lower availability and high production costs. The use of extremozymes in biosensing does not parallel their success in industrial applications. In recent years, the “collection” of recognition elements was enriched by extremozymes with interesting selectivity and by thermostable chimeras. The perspectives for biosensor development are exciting, considering also the progress in genetic editing for the oriented immobilization of enzymes, efficient folding, and better electron transport. Stability, production costs and immobilization at sensing interfaces must be improved to encourage wider applications of extremozymes in biosensors.
]]>Biosensors doi: 10.3390/bios14030144
Authors: Yinglu Li Shuhua Wei Enyi Xiong Jiawei Hu Xufang Zhang Yanrong Wang Jing Zhang Jiang Yan Zhaohao Zhang Huaxiang Yin Qingzhu Zhang
Silicon nanowire field effect (SiNW-FET) biosensors have been successfully used in the detection of nucleic acids, proteins and other molecules owing to their advantages of ultra-high sensitivity, high specificity, and label-free and immediate response. However, the presence of the Debye shielding effect in semiconductor devices severely reduces their detection sensitivity. In this paper, a three-dimensional stacked silicon nanosheet FET (3D-SiNS-FET) biosensor was studied for the high-sensitivity detection of nucleic acids. Based on the mainstream Gate-All-Around (GAA) fenestration process, a three-dimensional stacked structure with an 8 nm cavity spacing was designed and prepared, allowing modification of probe molecules within the stacked cavities. Furthermore, the advantage of the three-dimensional space can realize the upper and lower complementary detection, which can overcome the Debye shielding effect and realize high-sensitivity Point of Care Testing (POCT) at high ionic strength. The experimental results show that the minimum detection limit for 12-base DNA (4 nM) at 1 × PBS is less than 10 zM, and at a high concentration of 1 µM DNA, the sensitivity of the 3D-SiNS-FET is approximately 10 times higher than that of the planar devices. This indicates that our device provides distinct advantages for detection, showing promise for future biosensor applications in clinical settings.
]]>Biosensors doi: 10.3390/bios14030141
Authors: Yeong-Ung Kim Won-Ju Cho
Bovine serum albumin (BSA) is commonly incorporated in vaccines to improve stability. However, owing to potential allergic reactions in humans, the World Health Organization (WHO) mandates strict adherence to a BSA limit (≤50 ng/vaccine). BSA detection with conventional techniques is time-consuming and requires specialized equipment. Efficient alternatives such as the ion-sensitive field-effect transistor (ISFET), despite rapid detection, affordability, and portability, do not detect BSA at low concentrations because of inherent sensitivity limitations. This study proposes a silicon-on-insulator (SOI) substrate-based dual-gate (DG) ISFET platform to overcome these limitations. The capacitive coupling DG structure significantly enhances sensitivity without requiring external circuits, owing to its inherent amplification effect. The extended-gate (EG) structure separates the transducer unit for electrical signal processing from the sensing unit for biological detection, preventing chemical damage to the transducer, accommodating a variety of biological analytes, and affording easy replaceability. Vapor-phase surface treatment with (3-Aminopropyl) triethoxysilane (APTES) and the incorporation of a SnO2 sensing membrane ensure high BSA detection efficiency and sensitivity (144.19 mV/log [BSA]). This DG-FET-based biosensor possesses a simple structure and detects BSA at low concentrations rapidly. Envisioned as an effective on-site diagnostic tool for various analytes including BSA, this platform addresses prior limitations in biosensing and shows promise for practical applications.
]]>Biosensors doi: 10.3390/bios14030140
Authors: Suyeon Ahn Namju Kim Yonghyun Choi Jiwon Kim Hyeryun Hwang Cholong Kim Hee-Young Lee Seungyoun Kim Jin Su Kim Hyun Ho Lee Jonghoon Choi
The escalating utilization of plastics in daily life has resulted in pervasive environmental pollution and consequent health hazards. The challenge of detecting and capturing microplastics, which are imperceptible to the naked eye, is exacerbated by their diminutive size, hydrophobic surface properties, and capacity to absorb organic compounds. This study focuses on the application of peptides, constituted of specific amino acid sequences, and microneedles for the rapid and selective identification of microplastics. Peptides, due to their smaller size and greater environmental stability compared with antibodies, emerge as a potent solution to overcome the limitations inherent in existing detection methodologies. To immobilize peptides onto microneedles, this study employed microneedles embedded with gold nanorods, augmenting them with sulfhydryl (SH) groups at the peptides’ termini. The sensor developed through this methodology exhibited efficient peptide binding to the microneedle tips, thereby facilitating the capture of microplastics. Raman spectroscopy was employed for the detection of microplastics, with the results demonstrating successful attachment to the microneedles. This novel approach not only facilitates localized analysis but also presents a viable strategy for the detection of microplastics across diverse environmental settings.
]]>Biosensors doi: 10.3390/bios14030139
Authors: Yanju Chen Yuanyuan Zhu Cheng Peng Xiaofu Wang Jian Wu Huan Chen Junfeng Xu
Nucleic acid quantification, allowing us to accurately know the copy number of target nucleic acids, is significant for diagnosis, food safety, agricultural production, and environmental protection. However, current digital quantification methods require expensive instruments or complicated microfluidic chips, making it difficult to popularize in the point-of-care detection. Paper is an inexpensive and readily available material. In this study, we propose a simple and cost-effective paper membrane-based digital loop-mediated isothermal amplification (LAMP) method for nucleic acid quantification. In the presence of DNA fluorescence dyes, the high background signals will cover up the amplicons-formed bright spots. To reduce the background fluorescence signals, a quencher-fluorophore duplex was introduced in LAMP primers to replace non-specific fluorescence dyes. After that, the amplicons-formed spots on the paper membrane can be observed; thus, the target DNA can be quantified by counting the spots. Take Vibrio parahaemolyticus DNA detection as an instance, a good linear relationship is obtained between the light spots and the copy numbers of DNA. The paper membrane-based digital LAMP detection can detect 100 copies target DNA per reaction within 30 min. Overall, the proposed nucleic acid quantification method has the advantages of a simple workflow, short sample-in and answer-out time, low cost, and high signal-to-noise, which is promising for application in resourced limited areas.
]]>Biosensors doi: 10.3390/bios14030138
Authors: Asta Kausaite-Minkstimiene Algimantas Kaminskas Galina Gayda Almira Ramanaviciene
This paper describes the study of an amperometric glucose biosensor based on an enzymatic biofuel cell consisting of a bioanode and a biocathode modified with the same enzyme—glucose oxidase (GOx). A graphite rod electrode (GRE) was electrochemically modified with a layer of Prussian blue (PB) nanoparticles embedded in a poly(pyrrole-2-carboxylic acid) (PPCA) shell, and an additional layer of PPCA and was used as the cathode. A GRE modified with a nanocomposite composed of poly(1,10-phenanthroline-5,6-dione) (PPD) and gold nanoparticles (AuNPs) entrapped in a PPCA shell was used as an anode. Both electrodes were modified with GOx by covalently bonding the enzyme to the carboxyl groups of PPCA. The developed biosensor exhibited a wide linear range of 0.15–124.00 mM with an R2 of 0.9998 and a sensitivity of 0.16 μA/mM. The limit of detection (LOD) and quantification (LOQ) were found to be 0.07 and 0.23 mM, respectively. The biosensor demonstrated exceptional selectivity to glucose and operational stability throughout 35 days, as well as good reproducibility, repeatability, and anti-interference ability towards common interfering substances. The studies on human serum demonstrate the ability of the newly designed biosensor to determine glucose in complex real samples at clinically relevant concentrations.
]]>Biosensors doi: 10.3390/bios14030137
Authors: Yun Ho Jeong Mina Kwon Sangsoo Shin Jaegeun Lee Ki Su Kim
Carbon nanotubes (CNTs) have been regarded as emerging materials in various applications. However, the range of biomedical applications is limited due to the aggregation and potential toxicity of powder-type CNTs. To overcome these issues, techniques to assemble them into various macroscopic structures, such as one-dimensional fibers, two-dimensional films, and three-dimensional aerogels, have been developed. Among them, carbon nanotube fiber (CNTF) is a one-dimensional aggregate of CNTs, which can be used to solve the potential toxicity problem of individual CNTs. Furthermore, since it has unique properties due to the one-dimensional nature of CNTs, CNTF has beneficial potential for biomedical applications. This review summarizes the biomedical applications using CNTF, such as the detection of biomolecules or signals for biosensors, strain sensors for wearable healthcare devices, and tissue engineering for regenerating human tissues. In addition, by considering the challenges and perspectives of CNTF for biomedical applications, the feasibility of CNTF in biomedical applications is discussed.
]]>Biosensors doi: 10.3390/bios14030136
Authors: Sylwia Karoń Marcin Drozd Elżbieta Malinowska
Protein-based microarrays are important tools for high-throughput medical diagnostics, offering versatile platforms for multiplex immunodetection. However, challenges arise in protein microarrays due to the heterogeneous nature of proteins and, thus, differences in their immobilization conditions. This article advocates DNA-directed immobilization (DDI) as a solution, emphasizing its rapid and cost-effective fabrication of biosensing platforms. Thiolated single-stranded DNA and its analogues, such as ZNA® and PNA probes, were used to immobilize model proteins (anti-CRP antibodies and SARS-CoV nucleoprotein). The study explores factors influencing DDI-based immunosensor performance, including the purity of protein-DNA conjugates and the stability of their duplexes with DNA and analogues. It also provides insight into backfilling agent type and probe surface density. The research reveals that single-component monolayers lack protection against protein adsorption, while mixing the probes with long-chain ligands may hinder DNA-protein conjugate anchoring. Conventional DNA probes offer slightly higher surface density, while ZNA® probes exhibit better binding efficiency. Despite no enhanced stability in different ionic strength media, the cost-effectiveness of DNA probes led to their preference. The findings contribute to advancing microarray technology, paving the way for new generations of DDI-based multiplex platforms for rapid and robust diagnostics.
]]>Biosensors doi: 10.3390/bios14030135
Authors: Hui Chen Zhiyuan Zhuang Naihan Xu Ying Feng Kaixin Fang Chunyan Tan Ying Tan
The ongoing COVID-19 pandemic, driven by persistent SARS-CoV-2 transmission, threatens human health worldwide, underscoring the urgent need for an efficient, low-cost, rapid SARS-CoV-2 detection method. Herein, we developed a point-of-care SARS-CoV-2 detection method incorporating recombinase polymerase amplification (RPA) and DNA–protein crosslinking chemiluminescence (DPCL) (RPADPCL). RPADPCL involves the crosslinking of biotinylated double-stranded RPA DNA products with horseradish peroxidase (HRP)-labeled streptavidin (SA-HRP). Modified products are captured using SA-labeled magnetic beads, and then analyzed using a chemiluminescence detector and smartphone after the addition of a chemiluminescent substrate. Under optimal conditions, the RPADPCL limit of detection (LOD) was observed to be 6 copies (within the linear detection range of 1–300 copies) for a plasmid containing the SARS-CoV-2 N gene and 15 copies (within the linear range of 10–500 copies) for in vitro transcribed (IVT) SARS-CoV-2 RNA. The proposed method is convenient, specific, visually intuitive, easy to use, and does not require external excitation. The effective RPADPCL detection of SARS-CoV-2 in complex matrix systems was verified by testing simulated clinical samples containing 10% human saliva or a virus transfer medium (VTM) spiked with a plasmid containing a SARS-CoV-2 N gene sequence or SARS-CoV-2 IVT RNA. Consequently, this method has great potential for detecting targets in clinical samples.
]]>Biosensors doi: 10.3390/bios14030134
Authors: Natalija German Anton Popov Almira Ramanaviciene
Two types of low-cost reagentless electrochemical glucose biosensors based on graphite rod (GR) electrodes were developed. The electrodes modified with electrochemically synthesized platinum nanostructures (PtNS), 1,10-phenanthroline-5,6-dione (PD), glucose oxidase (GOx) without and with a polypyrrole (Ppy) layer—(i) GR/PtNS/PD/GOx and (ii) GR/PtNS/PD/GOx/Ppy, respectively, were prepared and tested. Glucose biosensors based on GR/PtNS/PD/GOx and GR/PtNS/PD/GOx/Ppy electrodes were characterized by the sensitivity of 10.1 and 5.31 μA/(mM cm2), linear range (LR) up to 16.5 and 39.0 mM, limit of detection (LOD) of 0.198 and 0.561 mM, good reproducibility, and storage stability. The developed glucose biosensors based on GR/PtNS/PD/GOx/Ppy electrodes showed exceptional resistance to interfering compounds and proved to be highly efficient for the determination of glucose levels in blood serum.
]]>Biosensors doi: 10.3390/bios14030133
Authors: Hyunjun Park Gayoung Kim Woochang Kim Eugene Park Joohyung Park Jinsung Park
Thiabendazole (TBZ) is a benzimidazole; owing to its potent antimicrobial properties, TBZ is extensively employed in agriculture as a fungicide and pesticide. However, TBZ poses environmental risks, and excessive exposure to TBZ through various leakage pathways can cause adverse effects in humans. Therefore, a method must be developed for early and sensitive detection of TBZ over a range of concentrations, considering both human and environmental perspectives. In this study, we used silver nanopillar structures (SNPis) and Au@Ag bimetallic nanoparticles (BNPs) to fabricate a BNP@SNPi substrate. This substrate exhibited a broad reaction surface with significantly enhanced surface-enhanced Raman scattering hotspots, demonstrating excellent Raman performance, along with high reproducibility, sensitivity, and selectivity for TBZ detection. Ultimately, the BNP@SNPi substrate successfully detected TBZ across a wide concentration range in samples of tap water, drinking water, juice, and human serum, with respective limits of detection of 146.5, 245.5, 195.6, and 219.4 pM. This study highlights BNP@SNPi as a promising sensor platform for TBZ detection in diverse environments and contributes to environmental monitoring and bioanalytical studies.
]]>Biosensors doi: 10.3390/bios14030132
Authors: Michal Katan Ori Pearl Alon Tzroya Hamootal Duadi Dror Fixler
Traditional methods for measuring blood oxygen use multiple wavelengths, which produce an intrinsic error due to ratiometric measurements. These methods assume that the absorption changes with the wavelength, but in fact the scattering changes as well and cannot be neglected. We found that if one measures in a specific angle around a cylindrical tissue, called the iso-pathlength (IPL) point, the reemitted light intensity is unaffected by the tissue’s scattering. Therefore, the absorption can be isolated from the scattering, which allows the extraction of the subject’s oxygen saturation. In this work, we designed an optical biosensor for reading the light intensity reemitted from the tissue, using a single light source and multiple photodetectors (PDs), with one of them in the IPL point’s location. Using this bio-device, we developed a methodology to extract the arterial oxygen saturation using a single wavelength light source. We proved this method is not dependent on the light source and is applicable to different measurement locations on the body, with an error of 0.5%. Moreover, we tested thirty-eight males and females with the biosensor under normal conditions. Finally, we show the results of measuring subjects in a hypoxic chamber that simulates extreme conditions with low oxygen.
]]>Biosensors doi: 10.3390/bios14030131
Authors: Deniz Mevlevioğlu Sabin Tabirca David Murphy
Virtual Reality Exposure Therapy is a method of cognitive behavioural therapy that aids in the treatment of anxiety disorders by making therapy practical and cost-efficient. It also allows for the seamless tailoring of the therapy by using objective, continuous feedback. This feedback can be obtained using biosensors to collect physiological information such as heart rate, electrodermal activity and frontal brain activity. As part of developing our objective feedback framework, we developed a Virtual Reality adaptation of the well-established emotional Stroop Colour–Word Task. We used this adaptation to differentiate three distinct levels of anxiety: no anxiety, mild anxiety and severe anxiety. We tested our environment on twenty-nine participants between the ages of eighteen and sixty-five. After analysing and validating this environment, we used it to create a dataset for further machine-learning classification of the assigned anxiety levels. To apply this information in real-time, all of our information was processed within Virtual Reality. Our Convolutional Neural Network was able to differentiate the anxiety levels with a 75% accuracy using leave-one-out cross-validation. This shows that our system can accurately differentiate between different anxiety levels.
]]>Biosensors doi: 10.3390/bios14030130
Authors: Juiena Hasan Sangho Bok
The increasing demand for rapid, cost-effective, and reliable diagnostic tools in personalized and point-of-care medicine is driving scientists to enhance existing technology platforms and develop new methods for detecting and measuring clinically significant biomarkers. Humanity is confronted with growing risks from emerging and recurring infectious diseases, including the influenza virus, dengue virus (DENV), human immunodeficiency virus (HIV), Ebola virus, tuberculosis, cholera, and, most notably, SARS coronavirus-2 (SARS-CoV-2; COVID-19), among others. Timely diagnosis of infections and effective disease control have always been of paramount importance. Plasmonic-based biosensing holds the potential to address the threat posed by infectious diseases by enabling prompt disease monitoring. In recent years, numerous plasmonic platforms have risen to the challenge of offering on-site strategies to complement traditional diagnostic methods like polymerase chain reaction (PCR) and enzyme-linked immunosorbent assays (ELISA). Disease detection can be accomplished through the utilization of diverse plasmonic phenomena, such as propagating surface plasmon resonance (SPR), localized SPR (LSPR), surface-enhanced Raman scattering (SERS), surface-enhanced fluorescence (SEF), surface-enhanced infrared absorption spectroscopy, and plasmonic fluorescence sensors. This review focuses on diagnostic methods employing plasmonic fluorescence sensors, highlighting their pivotal role in swift disease detection with remarkable sensitivity. It underscores the necessity for continued research to expand the scope and capabilities of plasmonic fluorescence sensors in the field of diagnostics.
]]>Biosensors doi: 10.3390/bios14030129
Authors: Yugyung Jung Seonjong Kim Min-Gon Kim Young-Eun Lee Myung-Geun Shin Sung Yang
Vancomycin (VAN) is an effective antibiotic against Gram-positive bacteria and the first-line therapy to prevent and treat methicillin-resistant Staphylococcus aureus (MRSA) and severe infections. However, low concentrations of VAN can result in resistant strains. High doses of VAN can cause nephrotoxicity and ototoxicity; thus, VAN is a representative drug for which drug monitoring is recommended. Several methods have been proposed to detect VAN. Among them, lateral flow immunoassays (LFIAs) have advantages, such as simple and user-friendly operation, low sample volume requirement, and cost effectiveness. In this study, we developed an LFIA capable of rapid on-site detection such that the VAN concentration in plasma could be monitored within 20 min by a one-step detection process using whole blood without plasma separation. VAN can be detected in whole blood over a wide range of concentrations (20−10,000 ng/mL), and the LFIA reported here has a detection limit of 18 ng/mL. The applicability of the developed LFIA compared to the results of measuring VAN with a commercial enzyme-linked immunosorbent assay kit showed a satisfactory correlation (Spearman’s rho, ρ = 0.891). Therefore, the developed LFIA enables rapid and wide-range VAN detection in whole blood and can aid in drug monitoring to evaluate patients’ responses to treatment.
]]>Biosensors doi: 10.3390/bios14030128
Authors: Bruno Dias Camargo Mateus Cassaboni Stracke Heloisa Bruna Soligo Sanchuki Viviane Klassen de Oliveira Hellen Cristina Ancelmo Dayanne Mozaner Bordin Fabricio Klerynton Marchini Emilson Ribeiro Viana Lucas Blanes
This work presents a low-cost transcription loop-mediated isothermal amplification (RT-LAMP) instrument for nucleic acid detection, employing an Arduino Nano microcontroller. The cooling system includes customized printed circuit boards (PCBs) that serve as electrical resistors and incorporate fans. An aluminum block is designed to accommodate eight vials. The system also includes two PCB heaters—one for sample heating and the other for vial lid heating to prevent condensation. The color detection system comprises a TCS3200 color 8-sensor array coupled to one side of the aluminum heater body and a white 8-LED array coupled to the other side, controlled by two Multiplexer/Demultiplexer devices. LED light passes through the sample, reaching the color sensor and conveying color information crucial for detection. The top board is maintained at 110 ± 2 °C, while the bottom board is held at 65 ± 0.5 °C throughout the RT-LAMP assay. Validation tests successfully demonstrated the efficacy of the colorimetric RT-LAMP reactions using SARS-CoV-2 RNA amplification as a sample viability test, achieving 100% sensitivity and 97.3% specificity with 66 clinical samples. Our instrument offers a cost-effective (USD 100) solution with automated result interpretation and superior sensitivity compared to visual inspection. While the prototype was tested with SARS-CoV-2 RNA samples, its versatility extends to detecting other pathogens using alternative primers, showcasing its potential for broader applications in biosensing.
]]>Biosensors doi: 10.3390/bios14030127
Authors: Alfa Rossi Yushun Zeng Mojtaba Rahimi Taeyoon Son Michael J. Heiferman Chen Gong Xin Sun Mohammad Soleimani Ali R. Djalilian Mark S. Humayun Qifa Zhou Xincheng Yao
Ultrasound A-scan is an important tool for quantitative assessment of ocular lesions. However, its usability is limited by the difficulty of accurately localizing the ultrasound probe to a lesion of interest. In this study, a transparent LiNbO3 single crystal ultrasound transducer was fabricated, and integrated with a widefield fundus camera to guide the ultrasound local position. The electrical impedance, phase spectrum, pulse-echo performance, and optical transmission spectrum of the ultrasound transducer were validated. The novel fundus camera-guided ultrasound probe was tested for in vivo measurement of rat eyes. Anterior and posterior segments of the rat eye could be unambiguously differentiated with the fundus photography-guided ultrasound measurement. A model eye was also used to verify the imaging performance of the prototype device in the human eye. The prototype shows the potential of being used in the clinic to accurately measure the thickness and echogenicity of ocular lesions in vivo.
]]>Biosensors doi: 10.3390/bios14030126
Authors: Călin Trif Dorin Harpaz Evgeni Eltzov Yardnapar Parcharoen Chiravoot Pechyen Robert S. Marks
An increasing number of countries have started to decriminalize or legalize the consumption of cannabis for recreational and medical purposes. The active ingredients in cannabis, termed cannabinoids, affect multiple functions in the human body, including coordination, motor skills, memory, response time to external stimuli, and even judgment. Cannabinoids are a unique class of terpeno-phenolic compounds, with 120 molecules discovered so far. There are certain situations when people under the influence of cannabis may be a risk to themselves or the public safety. Over the past two decades, there has been a growing research interest in detecting cannabinoids from various biological matrices. There is a need to develop a rapid, accurate, and reliable method of detecting cannabinoids in oral fluid as it can reveal the recent intake in comparison with urine specimens, which only show a history of consumption. Significant improvements are continuously made in the analytical formats of various technologies, mainly concerning improving their sensitivity, miniaturization, and making them more user-friendly. Additionally, sample collection and pretreatment have been extensively studied, and specific devices for collecting oral fluid specimens have been perfected to allow rapid and effective sample collection. This review presents the recent findings regarding the use of oral fluid specimens as the preferred biological matrix for cannabinoid detection in a point-of-care biosensor diagnostic device. A critical review is presented, discussing the findings from a collection of review and research articles, as well as publicly available data from companies that manufacture oral fluid screening devices. Firstly, the various conventional methods used to detect cannabinoids in biological matrices are presented. Secondly, the detection of cannabinoids using point-of-care biosensors is discussed, emphasizing oral fluid specimens. This review presents the current pressing technological challenges and highlights the gaps where new technological solutions can be implemented.
]]>Biosensors doi: 10.3390/bios14030125
Authors: Xiaoxuan Xu Yimei Zuo Shu Chen Amir Hatami Hui Gu
Neurochemicals, crucial for nervous system function, influence vital bodily processes and their fluctuations are linked to neurodegenerative diseases and mental health conditions. Monitoring these compounds is pivotal, yet the intricate nature of the central nervous system poses challenges. Researchers have devised methods, notably electrochemical sensing with micro-nanoscale electrodes, offering high-resolution monitoring despite low concentrations and rapid changes. Implantable sensors enable precise detection in brain tissues with minimal damage, while microdialysis-coupled platforms allow in vivo sampling and subsequent in vitro analysis, addressing the selectivity issues seen in other methods. While lacking temporal resolution, techniques like HPLC and CE complement electrochemical sensing’s selectivity, particularly for structurally similar neurochemicals. This review covers essential neurochemicals and explores miniaturized electrochemical sensors for brain analysis, emphasizing microdialysis integration. It discusses the pros and cons of these techniques, forecasting electrochemical sensing’s future in neuroscience research. Overall, this comprehensive review outlines the evolution, strengths, and potential applications of electrochemical sensing in the study of neurochemicals, offering insights into future advancements in the field.
]]>Biosensors doi: 10.3390/bios14030124
Authors: Seonghwan Shin Sangwon Kim Wonyeong Choi Jeonghyeon Do Jongmin Son Kihyun Kim Sungkey Jang Jeong-Soo Lee
The sensing responses of SARS-CoV-2 spike protein using top-down-fabricated Si-based electrolyte-gated transistors (EGTs) have been investigated. An aptamer was employed as a receptor for the SARS-CoV-2 spike protein. The EGT demonstrated excellent intrinsic characteristics and higher sensitivity in the subthreshold regime compared to the linear regime. The limit of detection (LOD) was achieved as low as 0.94 pg/mL and 20 pg/mL for the current and voltage sensitivity, respectively. To analyze the sensing responses of EGT in detecting the aptamer–SARS-CoV-2 spike protein conjugate, a lumped-capacitive model with the presence of an effective dipole potential and an effective capacitance of the functionalized layer component was employed. The aptamer-functionalized EGT showed high sensitivity even in 10 mM phosphate-buffered saline (PBS) solution. These results suggest that Si-based EGTs are a highly promising method for detecting SARS-CoV-2 spike proteins.
]]>Biosensors doi: 10.3390/bios14030123
Authors: Cristiana Domingues Rui Meirinho Rodolfo G. Rodrigues Ana Margarida Fortes Virginia Chu João Pedro Conde
Viticulture and associated products are an important part of the economy in many countries. However, biotic and abiotic stresses impact negatively the production of grapes and wine. Climate change is in many aspects increasing both these stresses. Routine sample retrievals and analysis tend to be time-consuming and require expensive equipment and skilled personnel to operate. These challenges could be overcome through the development of a miniaturized analytic device for early detection of grapevine stresses in the field. Abscisic acid is involved in several plant processes, including the onset of fruit ripening and tolerance mechanisms against drought stress. This hormone can be detected through a competitive immunoassay and is found in plants in concentrations up to 10−1 mg/mL. A microfluidic platform is developed in this work which can detect a minimum of 10−11 mg/mL of abscisic acid in buffer. Grape samples were tested using the microfluidic system alongside benchmark techniques such as high-performance liquid chromatography. The microfluidic system could detect the increase to 10−5 mg/mL of abscisic acid present in real berry samples at the veraison stage of ripening.
]]>Biosensors doi: 10.3390/bios14030122
Authors: Afagh Rastmanesh Jayanta S. Boruah Min-Seok Lee Seungkyung Park
Bioaerosols are small airborne particles composed of microbiological fragments, including bacteria, viruses, fungi, pollens, and/or by-products of cells, which may be viable or non-viable wherever applicable. Exposure to these agents can cause a variety of health issues, such as allergic and infectious diseases, neurological disorders, and cancer. Therefore, detecting and identifying bioaerosols is crucial, and bioaerosol sampling is a key step in any bioaerosol investigation. This review provides an overview of the current bioaerosol sampling methods, both passive and active, as well as their applications and limitations for rapid on-site monitoring. The challenges and trends for detecting airborne microorganisms using molecular and immunological methods are also discussed, along with a summary and outlook for the development of prompt monitoring technologies.
]]>Biosensors doi: 10.3390/bios14030121
Authors: Chuanxiang Zhang Song Li Lingxiao Tang Shuo Li Changchun Hu Dan Zhang Long Chao Xueying Liu Yimin Tan Yan Deng
Development of an efficient technique for accurate and sensitive dibutyl phthalate (DBP) determination is crucial for food safety and environment protection. An ultrasensitive molecularly imprinted polymers (MIP) voltammetric sensor was herein engineered for the specific determination of DBP using poly-l-lysine/poly(3,4-ethylenedioxythiophene)/porous graphene nanocomposite (PLL/PEDOT−PG) and poly(o-phenylenediamine)-imprinted film as a label-free and sensing platform. Fabrication of PEDOT−PG nanocomposites was achieved through a simple liquid–liquid interfacial polymerization. Subsequently, poly-l-lysine (PLL) functionalization was employed to enhance the dispersibility and stability of the prepared PEDOT−PG, as well as promote its adhesion on the sensor surface. In the presence of DBP, the imprinted poly(o-phenylenediamine) film was formed on the surface of PLL/PEDOT−PG. Investigation of the physical properties and electrochemical behavior of the MIP/PLL/PEDOT−PG indicates that the incorporation of PG into PEDOT, with PLL uniformly wrapping its surface, significantly enhanced conductivity, carrier mobility, stability, and provided a larger surface area for specific recognition sites. Under optimal experimental conditions, the electrochemical response exhibited a linear relationship with a logarithm of DBP concentration within the range of 1 fM to 5 µM, with the detection limit as low as 0.88 fM. The method demonstrated exceptional stability and repeatability and has been successfully applied to quantify DBP in plastic packaging materials.
]]>Biosensors doi: 10.3390/bios14030120
Authors: Dmitry Stoikov Alexey Ivanov Insiya Shafigullina Milena Gavrikova Pavel Padnya Igor Shiabiev Ivan Stoikov Gennady Evtugyn
A flow-through biosensor system for the determination of uric acid was developed on the platform of flow-through electrochemical cell manufactured by 3D printing from poly(lactic acid) and equipped with a modified screen-printed graphite electrode (SPE). Uricase was immobilized to the inner surface of a replaceable reactor chamber. Its working volume was reduced to 10 μL against a previously reported similar cell. SPE was modified independently of the enzyme reactor with carbon black, pillar[5]arene, poly(amidoamine) dendrimers based on the p-tert-butylthiacalix[4]arene (PAMAM-calix-dendrimers) platform and electropolymerized 3,7-bis(4-aminophenylamino) phenothiazin-5-ium chloride. Introduction of the PAMAM-calix-dendrimers into the electrode coating led to a fivefold increase in the redox currents of the electroactive polymer. It was found that higher generations of the PAMAM-calix-dendrimers led to a greater increase in the currents measured. Coatings consisted of products of the electropolymerization of the phenothiazine with implemented pillar[5]arene and PAMAM-calix-dendrimers showing high efficiency in the electrochemical reduction of hydrogen peroxide that was formed in the enzymatic oxidation of uric acid. The presence of PAMAM-calix-dendrimer G2 in the coating increased the redox signal related to the uric acid assay by more than 1.5 times. The biosensor system was successfully applied for the enzymatic determination of uric acid in chronoamperometric mode. The following optimal parameters for the chronoamperometric determination of uric acid in flow-through conditions were established: pH 8.0, flow rate 0.2 mL·min−1, 5 U of uricase per reactor. Under these conditions, the biosensor system made it possible to determine from 10 nM to 20 μM of uric acid with the limit of detection (LOD) of 4 nM. Glucose (up to 1 mM), dopamine (up to 0.5 mM), and ascorbic acid (up to 50 μM) did not affect the signal of the biosensor toward uric acid. The biosensor was tested on spiked artificial urine samples, and showed 101% recovery for tenfold diluted samples. The ease of assembly of the flow cell and the low cost of the replacement parts make for a promising future application of the biosensor system in routine clinical analyses.
]]>Biosensors doi: 10.3390/bios14030119
Authors: Alaleh Vaghef-Koodehi Blanca H. Lapizco-Encinas
Analyte migration order is a major aspect in all migration-based analytical separations methods. Presented here is the manipulation of the migration order of microparticles in an insulator-based electrokinetic separation. Three distinct particle mixtures were studied: a binary mixture of particles with similar electrical charge and different sizes, and two tertiary mixtures of particles of distinct sizes. Each one of the particle mixtures was separated twice, the first separation was performed under low voltage (linear electrokinetic regime) and the second separation was performed under high voltage (nonlinear electrokinetic regime). Linear electrophoresis, which discriminates particles by charge, is the dominant electrokinetic effect in the linear regime; while nonlinear electrophoresis, which discriminates particles by size and shape, is the dominant electrokinetic effect in the nonlinear regime. The separation results obtained with the three particle mixtures illustrated that particle elution order can be changed by switching from the linear electrokinetic regime to the nonlinear electrokinetic regime. Also, in all cases, better separation performances in terms of separation resolution (Rs) were obtained by employing the nonlinear electrokinetic regime allowing nonlinear electrophoresis to be the discriminatory electrokinetic mechanism. These findings could be applied to analyze complex samples containing bioparticles of interest within the micron size range. This is the first report where particle elution order is altered in an iEK system.
]]>Biosensors doi: 10.3390/bios14030118
Authors: Brian Sang Haoran Wen Gregory Junek Wendy Neveu Lorenzo Di Francesco Farrokh Ayazi
Wheezing is a critical indicator of various respiratory conditions, including asthma and chronic obstructive pulmonary disease (COPD). Current diagnosis relies on subjective lung auscultation by physicians. Enabling this capability via a low-profile, objective wearable device for remote patient monitoring (RPM) could offer pre-emptive, accurate respiratory data to patients. With this goal as our aim, we used a low-profile accelerometer-based wearable system that utilizes deep learning to objectively detect wheezing along with respiration rate using a single sensor. The miniature patch consists of a sensitive wideband MEMS accelerometer and low-noise CMOS interface electronics on a small board, which was then placed on nine conventional lung auscultation sites on the patient’s chest walls to capture the pulmonary-induced vibrations (PIVs). A deep learning model was developed and compared with a deterministic time–frequency method to objectively detect wheezing in the PIV signals using data captured from 52 diverse patients with respiratory diseases. The wearable accelerometer patch, paired with the deep learning model, demonstrated high fidelity in capturing and detecting respiratory wheezes and patterns across diverse and pertinent settings. It achieved accuracy, sensitivity, and specificity of 95%, 96%, and 93%, respectively, with an AUC of 0.99 on the test set—outperforming the deterministic time–frequency approach. Furthermore, the accelerometer patch outperforms the digital stethoscopes in sound analysis while offering immunity to ambient sounds, which not only enhances data quality and performance for computational wheeze detection by a significant margin but also provides a robust sensor solution that can quantify respiration patterns simultaneously.
]]>Biosensors doi: 10.3390/bios14030117
Authors: Yanping Sun Jianxin Ma Faisal Ahmad Yelan Xiao Jingyang Guan Tong Shu Xueji Zhang
Bimetallic coordination polymers (CPs) have two different metal ions as connecting nodes in their polymer structure. The synthesis methods of bimetallic CPs are mainly categorized into the one-pot method and post-synthesis modifications according to various needs. Compared with monometallic CPs, bimetallic CPs have synergistic effects and excellent properties, such as higher gas adsorption rate, more efficient catalytic properties, stronger luminescent properties, and more stable loading platforms, which have been widely applied in the fields of gas adsorption, catalysis, energy storage as well as conversion, and biosensing. In recent years, the study of bimetallic CPs synergized with cancer drugs and functional nanomaterials for the therapy of cancer has increasingly attracted the attention of scientists. This review presents the research progress of bimetallic CPs in biosensing and biomedicine in the last five years and provides a perspective for their future development.
]]>Biosensors doi: 10.3390/bios14030116
Authors: Neha Dubey Sudeshna Chandra
Electrospun nanofibers possess a large surface area and a three-dimensional porous network that makes them a perfect material for embedding functional nanoparticles for diverse applications. Herein, we report the trends in embedding upconversion nanoparticles (UCNPs) in polymeric nanofibers for making an advanced miniaturized (bio)analytical device. UCNPs have the benefits of several optical properties, like near-infrared excitation, anti-Stokes emission over a wide range from UV to NIR, narrow emission bands, an extended lifespan, and photostability. The luminescence of UCNPs can be regulated using different lanthanide elements and can be used for sensing and tracking physical processes in biological systems. We foresee that a UCNP-based nanofiber sensing platform will open opportunities in developing cost-effective, miniaturized, portable and user-friendly point-of-care sensing device for monitoring (bio)analytical processes. Major challenges in developing microfluidic (bio)analytical systems based on UCNPs@nanofibers have been reviewed and presented.
]]>Biosensors doi: 10.3390/bios14030115
Authors: Vishnu Sharma Hitomi Takamura Manish Biyani Ryo Honda
Wastewater-based epidemiology (WBE) is an effective and efficient tool for the early detection of infectious disease outbreaks in a community. However, currently available methods are laborious, costly, and time-consuming due to the low concentration of viruses and the presence of matrix chemicals in wastewater that may interfere with molecular analyses. In the present study, we designed a highly sensitive “Quick Poop (wastewater with fecal waste) Sensor” (termed, QPsor) using a joint approach of Nanotrap microbiome particles and RICCA (RNA Isothermal Co-Assisted and Coupled Amplification). Using QPsor, the WBE study showed a strong correlation with standard PEG concentrations and the qPCR technique. Using a closed format for a paper-based lateral flow assay, we were able to demonstrate the potential of our assay as a real-time, point-of-care test by detecting the heat-inactivated SARS-CoV-2 virus in wastewater at concentrations of 100 copies/mL and within one hour. As a proof-of-concept demonstration, we analyzed the presence of viral RNA of the SARS-CoV-2 virus and PMMoV in raw wastewater samples from wastewater treatment plants on-site and within 60 min. The results show that the QPsor method can be an effective tool for disease outbreak detection by combining an AI-enabled case detection model with real-time on-site viral RNA extraction and amplification, especially in the absence of intensive clinical laboratory facilities. The lab-free, lab-quality test capabilities of QPsor for viral prevalence and transmission in the community can contribute to the efficient management of pandemic situations.
]]>Biosensors doi: 10.3390/bios14030114
Authors: Shiyuan Gao Tiegang Xu Lei Wu Xiaoyue Zhu Xuefeng Wang Ying Chen Gang Li Xinxin Li
In a chamber-based digital PCR (dPCR) chip fabricated with polydimethylsiloxane (PDMS), bubble generation in the chambers at high temperatures is a critical issue. Here, we found that the main reason for bubble formation in PDMS chips is the too-high saturated vapor pressure of water at an elevated temperature. The bubbles should be completely prevented by reducing the initial pressure of the system to under 13.6 kPa to eliminate the effects of increased-pressure water vapor. Then, a cavity was designed and fabricated above the PCR reaction layer, and Parylene C was used as a shell covering the chip. The cavity was used for the negative generator in sample loading, PDMS degassing, PCR solution degassing in the digitization process and water storage in the thermal reaction process. The analysis was confirmed and finally achieved a desirable bubble-free, fast-digitization, valve-free and no-tubing connection dPCR.
]]>Biosensors doi: 10.3390/bios14030113
Authors: Ki Sung Park Anna Choi Tae-In Park Seung Pil Pack
SARS-CoV-2, the virus responsible for the COVID-19 pandemic, has spurred the urgent need for practical diagnostics with high sensitivity and selectivity. Although advanced diagnostic tools have emerged to efficiently control pandemics, they still have costly limitations owing to their reliance on antibodies or enzymes and require high-tech equipment. Therefore, there is still a need to develop rapid and low-cost diagnostics with high sensitivity and selectivity. In this study, we generated aptamer display particles (AdP), enabling easy fabrication of a SARS-CoV-2 detection matrix through particle PCR, and applied it to diagnosis using fluorometric and colorimetric assays. We designed two AdPs, C1-AdP and C4-AdP, displayed with SpS1-C1 and SpS1-C4 aptamers, respectively, and showed their high binding ability against SARS-CoV-2 spike protein with a concentration-dependent fluorescence increase. This enabled detection even at low concentrations (0.5 nM). To validate its use as a diagnostic tool for SARS-CoV-2, we designed a sandwich-type assay using two AdPs and high-quality aptamers targeting SARS-CoV-2 pseudoviruses. The fluorometric assay achieved a detection limit of 3.9 × 103 pseudoviruses/mL. The colorimetric assay using an amplification approach exhibited higher sensitivity, with a detection limit of 1 × 101 pseudoviruses/mL, and a broad range of over four orders of magnitude was observed.
]]>Biosensors doi: 10.3390/bios14030112
Authors: Shaoxi Wang Zhexin Zhang Xun Ma Yuanbo Yue Kemu Li Yingqi Meng Yupan Wu
The rotation of cells is of significant importance in various applications including bioimaging, biophysical analysis and microsurgery. Current methods usually require complicated fabrication processes. Herein, we proposed an induced charged electroosmosis (ICEO) based on a chip manipulation method for rotating cells. Under an AC electric field, symmetric ICEO flow microvortexes formed above the electrode surface can be used to trap and rotate cells. We have discussed the impact of ICEO and dielectrophoresis (DEP) under the experimental conditions. The capabilities of our method have been tested by investigating the precise rotation of yeast cells and K562 cells in a controllable manner. By adjusting the position of cells, the rotation direction can be changed based on the asymmetric ICEO microvortexes via applying a gate voltage to the gate electrode. Additionally, by applying a pulsed signal instead of a continuous signal, we can also precisely and flexibly rotate cells in a stepwise way. Our ICEO-based rotational manipulation method is an easy to use, biocompatible and low-cost technique, allowing rotation regardless of optical, magnetic or acoustic properties of the sample.
]]>Biosensors doi: 10.3390/bios14020111
Authors: Zhen Wang Xiao Wang Guijun Shu Meng Yin Shoushuang Huang Ming Yin
This article presents the design of a low-power, low-noise neural signal amplifier for neural recording. The structure reduces the current consumption of the amplifier through current scaling technology and lowers the input-referred noise of the amplifier by combining a source degeneration resistor and current reuse technologies. The amplifier was fabricated using a 0.18 μm CMOS MS RF G process. The results show the front-end amplifier exhibits a measured mid-band gain of 40 dB/46 dB and a bandwidth ranging from 0.54 Hz to 6.1 kHz; the amplifier’s input-referred noise was measured to be 3.1 μVrms, consuming a current of 3.8 μA at a supply voltage of 1.8 V, with a Noise Efficiency Factor (NEF) of 2.97. The single amplifier’s active silicon area is 0.082 mm2.
]]>Biosensors doi: 10.3390/bios14020110
Authors: Marta Jarczewska Marta Sokal Marcin Olszewski Elzbieta Malinowska
Lead poses severe effects on living organisms, and since Pb2+ ions tend to accumulate in different organs, it is crucial to monitor Pb2+ concentration in samples such as water and soil. One of the approaches is the utilization of biosensors combined with aptamer-based layers for the electrochemical detection of lead ions. Herein, we present the studies of applying miniaturized screen-printed transducers as solid surfaces to fabricate aptamer layers. As the research is the direct continuation of our previous studies regarding the use of gold disk electrodes, the working parameters of elaborated aptasensors were defined, including the range of linear response (10–100 nM), selectivity as well as stability, regeneration, and feasibility of application for the analysis of real samples. This was achieved using voltammetric techniques including cyclic and square-wave voltammetry in the presence of methylene blue redox indicator.
]]>Biosensors doi: 10.3390/bios14020109
Authors: Yeeun Na Chaehwan Kim Keunhoi Kim Tae Hyun Kim Soo Hyun Kwon Il-Suk Kang Young Woo Jung Tae Won Kim Deok-Ho Cho Jihwan An Jong-Kwon Lee Jongcheol Park
Reflection-type photoplethysmography (PPG) pulse sensors used in wearable smart watches, true wireless stereo, etc., have been recently considered a key component for monitoring biological signals such as heart rate, SPO3, and blood pressure. Typically, the optical front end (OFE) of these PPG sensors is heterogeneously configured and packaged with light sources and receiver chips. In this paper, a novel quarter-annulus photodetector (NQAPD) with identical inner and outer radii of curvature has been developed using a plasma dicing process to realize a ring-type OFE receiver, which maximizes manufacturing efficiency and increases the detector collection area by 36.7% compared to the rectangular PD. The fabricated NQAPD exhibits a high quantum efficiency of over 90% in the wavelength of 500 nm to 740 nm and the highest quantum efficiency of 95% with a responsivity of 0.41 A/W at the wavelength of 530 nm. Also, the NQAPD is shown to increase the SNR of the PPG signal by 5 to 7.6 dB compared to the eight rectangular PDs. Thus, reflective PPG sensors constructed with NQAPD can be applied to various wearable devices requiring low power consumption, high performance, and cost-effectiveness.
]]>Biosensors doi: 10.3390/bios14020108
Authors: Jael Abigail Medrano-Lopez Isaela Villalpando Ma Isabel Salazar Carlos Torres-Torres
Nanostructures have played a key role in the development of different techniques to attack severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Some applications include masks, vaccines, and biosensors. The latter are of great interest for detecting diseases since some of their features allowed us to find specific markers in secretion samples such as saliva, blood, and even tears. Herein, we highlight how hierarchical nanoparticles integrated into two or more low-dimensional materials present outstanding advantages that are attractive for photonic biosensing using their nanoscale functions. The potential of nanohybrids with their superlative mechanical characteristics together with their optical and optoelectronic properties is discussed. The progress in the scientific research focused on using nanoparticles for biosensing a variety of viruses has become a medical milestone in recent years, and has laid the groundwork for future disease treatments. This perspective analyzes the crucial information about the use of hierarchical nanostructures in biosensing for the prevention, treatment, and mitigation of SARS-CoV-2 effects.
]]>Biosensors doi: 10.3390/bios14020107
Authors: Mateo Gabriel Vasconez Martinez Eva I. Reihs Helene M. Stuetz Astrid Hafner Konstanze Brandauer Florian Selinger Patrick Schuller Neus Bastus Victor Puntes Johannes Frank Wolfgang Tomischko Martin Frauenlob Peter Ertl Christian Resch Gerald Bauer Guenter Povoden Mario Rothbauer
Due to advances in additive manufacturing and prototyping, affordable and rapid microfluidic sensor-integrated assays can be fabricated using additive manufacturing, xurography and electrode shadow masking to create versatile platform technologies aimed toward qualitative assessment of acute cytotoxic or cytolytic events using stand-alone biochip platforms in the context of environmental risk assessment. In the current study, we established a nasal mucosa biosensing platform using RPMI2650 mucosa cells inside a membrane-integrated impedance-sensing biochip using exclusively rapid prototyping technologies. In a final proof-of-concept, we applied this biosensing platform to create human cell models of nasal mucosa for monitoring the acute cytotoxic effect of zinc oxide reference nanoparticles. Our data generated with the biochip platform successfully monitored the acute toxicity and cytolytic activity of 6 mM zinc oxide nanoparticles, which was non-invasively monitored as a negative impedance slope on nasal epithelial models, demonstrating the feasibility of rapid prototyping technologies such as additive manufacturing and xurography for cell-based platform development.
]]>Biosensors doi: 10.3390/bios14020105
Authors: Xinmeng Zhang Cheng Zhou Jianxun Hou Gang Feng Zhourui Xu Yonghong Shao Chengbin Yang Gaixia Xu
Precise DNA quantification and nuclear imaging are pivotal for clinical testing, pathological diagnosis, and drug development. The detection and localization of mitochondrial DNA serve as crucial indicators of cellular health. We introduce a novel conjugated oligoelectrolyte (COE) molecule, COE-S3, featuring a planar backbone composed of three benzene rings and terminal side chains. This unique amphiphilic structure endows COE-S3 with exceptional water solubility, a high quantum yield of 0.79, and a significant fluorescence Stokes shift (λex = 366 nm, λem = 476 nm), alongside a specific fluorescence response to DNA. The fluorescence intensity correlates proportionally with DNA concentration. COE-S3 interacts with double-stranded DNA (dsDNA) through an intercalation binding mode, exhibiting a binding constant (K) of 1.32 × 106 M−1. Its amphiphilic nature and strong DNA affinity facilitate its localization within mitochondria in living cells and nuclei in apoptotic cells. Remarkably, within 30 min of COE-S3 staining, cell vitality can be discerned through real-time nuclear fluorescence imaging of apoptotic cells. COE-S3’s high DNA selectivity enables quantitative intracellular DNA analysis, providing insights into cell proliferation, differentiation, and growth. Our findings underscore COE-S3, with its strategically designed, shortened planar backbone, as a promising intercalative probe for DNA quantification and nuclear imaging.
]]>Biosensors doi: 10.3390/bios14020106
Authors: Tawfiq Alsulami Abdulhakeem Alzahrani
The needless use of tetracyclines (TCs) in foodstuffs is a huge health concern in low- and middle-income and Arab countries. Herein, a sensitive and faster monitoring system for H2O2 and TCs is proposed, utilizing the large surface-to-volume ratio of a non-spherical gold nanoparticle/black phosphorus nanocomposite (BP-nsAu NPs) for the first time. BP-nsAu NPs were synthesized through a single-step method that presented nanozymatic activity through 3,3′,5,5′-Tetramethylbenzidine (TMB) oxidation while H2O2 was present and obeyed the Michaelis–Menten equation. The nanozymatic activity of the BP-nsAu NPs was enhanced 12-fold and their detection time was decreased 83-fold compared to conventional nanozymatic reactions. The proposed method enabled us to quantify H2O2 with a limit of detection (LOD) value of 60 nM. Moreover, target-specific aptamer-conjugated BP-nsAu NPs helped us detect TCs with an LOD value of 90 nM. The present strategy provides a proficient route for low-level TC monitoring in real samples.
]]>Biosensors doi: 10.3390/bios14020104
Authors: Roselien Vercauteren Clémentine Gevers Jacques Mahillon Laurent A. Francis
The design of a porous silicon (PSi) biosensor is not often documented, but is of the upmost importance to optimize its performance. In this work, the motivation behind the design choices of a PSi-based optical biosensor for the indirect detection of bacteria via their lysis is detailed. The transducer, based on a PSi membrane, was characterized and models were built to simulate the analyte diffusion, depending on the porous nanostructures, and to optimize the optical properties. Once all performances and properties were analyzed and optimized, a theoretical response was calculated. The theoretical limit of detection was computed as 104 CFU/mL, based on the noise levels of the optical setup. The experimental response was measured using 106 CFU/mL of Bacillus cereus as model strain, lysed by bacteriophage-coded endolysins PlyB221. The obtained signal matched the expected response, demonstrating the validity of our design and models.
]]>Biosensors doi: 10.3390/bios14020103
Authors: Xuewen Lu Kangning Ding Zhiyuan Fang Yilei Liu Tianxing Ji Jian Sun Zhenling Zeng Limin He
Respiratory pathogens pose a huge threat to public health, especially the highly mutant RNA viruses. Therefore, reliable, on-site, rapid diagnosis of such pathogens is an urgent need. Traditional assays such as nucleic acid amplification tests (NAATs) have good sensitivity and specificity, but these assays require complex sample pre-treatment and a long test time. Herein, we present an on-site biosensor for rapid and multiplex detection of RNA pathogens. Samples with viruses are first lysed in a lysis buffer containing carrier RNA to release the target RNAs. Then, the lysate is used for amplification by one-step reverse transcription and single-direction isothermal strand displacement amplification (SDA). The yield single-strand DNAs (ssDNAs) are visually detected by a lateral flow biosensor. With a secondary signal amplification system, as low as 20 copies/μL of virus can be detected in this study. This assay avoids the process of nucleic acid purification, making it equipment-independent and easier to operate, so it is more suitable for on-site molecular diagnostic applications.
]]>Biosensors doi: 10.3390/bios14020102
Authors: Heekyung Park Jun Hui Park
Single-entity electrochemistry, which employs electrolysis during the collision of single particles on ultramicroelectrodes, has witnessed significant advancements in recent years, enabling the observation and characterization of individual particles. Information on a single aqueous droplet (e.g., size) can also be studied based on the redox species contained therein. Dopamine, a redox-active neurotransmitter, is usually present in intracellular vesicles. Similarly, in the current study, the electrochemical properties of neurotransmitters in submicron droplets were investigated. Because dopamine oxidation is accompanied by proton transfer, unique electrochemical properties of dopamine were observed in the droplet. We also investigated the electrochemical properties of the adsorbed droplets containing DA and the detection of oxidized dopamine by the recollision phenomenon.
]]>Biosensors doi: 10.3390/bios14020101
Authors: Kamila Malecka-Baturo Paulina Żółtowska Agnieszka Jackowska Katarzyna Kurzątkowska-Adaszyńska Iwona Grabowska
Here, we present the results of our the electrochemical aptasensing strategy for retinol binding protein-4 (RBP-4) detection based on a thiolated aptamer against RBP-4 and 6-mercaptohexanol (MCH) directly immobilized on a gold electrode surface. The most important parameters affecting the magnitude of the analytical signal generated were optimized: (i) the presence of magnesium ions in the immobilization and measurement buffer, (ii) the concentration of aptamer in the immobilization solution and (iii) its folding procedure. In this work, a systematic assessment of the electrochemical parameters related to the optimization of the sensing layer of the aptasensor was carried out (electron transfer coefficients (α), electron transfer rate constants (k0) and surface coverage of the thiolated aptamer probe (ΓApt)). Then, under the optimized conditions, the analytical response towards RBP-4 protein, in the presence of an Fe(CN)63−/4− redox couple in the supporting solution was assessed. The proposed electrochemical strategy allowed for RBP-4 detection in the concentration range between 100 and 1000 ng/mL with a limit of detection equal to 44 ng/mL based on electrochemical impedance spectroscopy (EIS). The specificity studies against other diabetes biomarkers, including vaspin and adiponectin, proved the selectivity of the proposed platform. These preliminary results will be used in the next step to miniaturize and test the sensor in real samples.
]]>Biosensors doi: 10.3390/bios14020100
Authors: Xiaohui Liu Hui Jiang Xuemei Wang
Cancers of unknown primary (CUP) exhibit significant cellular heterogeneity and malignancy, which poses significant challenges for diagnosis and treatment. Recent years have seen deeper insights into the imaging, pathology, and genetic characteristics of CUP, driven by interdisciplinary collaboration and the evolution of diagnostic and therapeutic strategies. However, due to their insidious onset, lack of evidence-based medicine, and limited clinical understanding, diagnosing and treating CUP remain a significant challenge. To inspire more creative and fantastic research, herein, we report and highlight recent advances in the diagnosis and therapeutic strategies of CUP. Specifically, we discuss advanced diagnostic technologies, including 12-deoxy-2-[fluorine-18]fluoro-D-glucose integrated with computed tomography (18F-FDG PET/CT) or 68Ga-FAPI (fibroblast activation protein inhibitor) PET/CT, liquid biopsy, molecular diagnostics, self-assembling nanotechnology, and artificial intelligence (AI). In particular, the discussion will extend to the effective treatment techniques currently available, such as targeted therapies, immunotherapies, and bio-nanotechnology-based therapeutics. Finally, a novel perspective on the challenges and directions for future CUP diagnostic and therapeutic strategies is discussed.
]]>Biosensors doi: 10.3390/bios14020099
Authors: Devi Taufiq Nurrohman Nan-Fu Chiu
Surface Plasmon Resonance (SPR) technology is known to be a powerful tool for studying biomolecular interactions because it offers real-time and label-free multiparameter analysis with high sensitivity. This article summarizes the results that have been obtained from the use of SPR technology in studying the dynamics of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) mutations. This paper will begin by introducing the working principle of SPR and the kinetic parameters of the sensorgram, which include the association rate constant (ka), dissociation rate constant (kd), equilibrium association constant (KA), and equilibrium dissociation constant (KD). At the end of the paper, we will summarize the kinetic data on the interaction between angiotensin-converting enzyme 2 (ACE2) and SARS-CoV-2 obtained from the results of SPR signal analysis. ACE2 is a material that mediates virus entry. Therefore, understanding the kinetic changes between ACE2 and SARS-CoV-2 caused by the mutation will provide beneficial information for drug discovery, vaccine development, and other therapeutic purposes.
]]>Biosensors doi: 10.3390/bios14020098
Authors: Rongwei Lei Hufsa Arain David Wang Janani Arunachalam Ramesh Saxena Chandra Mohan
The goal of this study is to develop a rapid diagnostic test for rheumatic disease and systemic lupus erythematosus (SLE) screening. A novel rapid vertical flow assay (VFA) was engineered and used to assay anti-nuclear (ANA) and anti-dsDNA (αDNA) autoantibodies from systemic lupus erythematosus (SLE) patients and healthy controls (HCs). Observer scores and absolute signal intensities from the VFA were validated via ELISA. The rapid point-of-care VFA test that was engineered demonstrated a limit of detection of 0.5 IU/mL for ANA and αDNA autoantibodies in human plasma with an inter-operator CV of 19% for ANA and 12% for αDNA. Storage stability was verified over a three-month period. When testing anti-dsDNA and ANA levels in SLE and HC serum samples, the duplex VFA revealed 95% sensitivity, 72% specificity and an 84% ROC AUC value in discriminating disease groups, comparable to the gold standard, ELISA. The rapid αDNA/ANA duplex VFA can potentially be used in primary care clinics for evaluating patients or at-risk subjects for rheumatic diseases and for planning follow-up testing. Given its low cost, ease, and rapid turnaround, it can also be used to assess SLE prevalence estimates.
]]>Biosensors doi: 10.3390/bios14020097
Authors: Le Thi Nhu Ngoc Young-Chul Lee
Ribonucleic acid (RNA) viruses are one of the major classes of pathogens that cause human diseases. The conventional method to detect RNA viruses is real-time quantitative reverse transcription polymerase chain reaction (qRT-PCR), but it has some limitations. It is expensive and time-consuming, with infrastructure and trained personnel requirements. Its high throughput requires sophisticated automation and large-scale infrastructure. Isothermal amplification methods have been explored as an alternative to address these challenges. These methods are rapid, user-friendly, low-cost, can be performed in less specialized settings, and are highly accurate for detecting RNA viruses. Microfluidic technology provides an ideal platform for performing virus diagnostic tests, including sample preparation, immunoassays, and nucleic acid-based assays. Among these techniques, nucleic acid isothermal amplification methods have been widely integrated with microfluidic platforms for RNA virus detection owing to their simplicity, sensitivity, selectivity, and short analysis time. This review summarizes some common isothermal amplification methods for RNA viruses. It also describes commercialized devices and kits that use isothermal amplification techniques for SARS-CoV-2 detection. Furthermore, the most recent applications of isothermal amplification-based microfluidic platforms for RNA virus detection are discussed in this article.
]]>Biosensors doi: 10.3390/bios14020096
Authors: Santhosh Kannan Chien-Chung Peng Hsiao-Mei Wu Yi-Chung Tung
Oxygen consumption has been used to evaluate various cellular activities. In addition, three-dimensional (3D) spheroids have been broadly exploited as advanced in vitro cell models for various biomedical studies due to their capability of mimicking 3D in vivo microenvironments and cell arrangements. However, monitoring the oxygen consumption of live 3D spheroids poses challenges because existing invasive methods cause structural and cell damage. In contrast, optical methods using fluorescence labeling and microscopy are non-invasive, but they suffer from technical limitations like high cost, tedious procedures, and poor signal-to-noise ratios. To address these challenges, we developed a microfluidic platform for uniform-sized spheroid formation, handling, and culture. The platform is further integrated with widefield frequency domain fluorescence lifetime imaging microscopy (FD-FLIM) to efficiently characterize the lifetime of an oxygen-sensitive dye filling the platform for oxygen consumption characterization. In the experiments, osteosarcoma (MG-63) cells are exploited as the spheroid model and for the oxygen consumption analysis. The results demonstrate the functionality of the developed approach and show the accurate characterization of the oxygen consumption of the spheroids in response to drug treatments. The developed approach possesses great potential to advance spheroid metabolism studies with single-spheroid resolution and high sensitivity.
]]>Biosensors doi: 10.3390/bios14020095
Authors: Wenhao Yuan Zeyu Zhao Shunsuke Kimura Kiyoshi Toko
A taste sensor employs various lipid/polymer membranes with specific physicochemical properties for taste classification and evaluation. However, phosphoric acid di(2-ethylhexyl) ester (PAEE), employed as one of the lipids for the taste sensors, exhibits insufficient selectivity for umami substances. The pH of sample solutions impacts the dissociation of lipids to influence the membrane potential, and the response to astringent substances makes accurate measurement of umami taste difficult. This study aims to develop a novel taste sensor for detecting umami substances like monosodium L-glutamate (MSG) through surface modification, i.e., a methodology previously applied to taste sensors for non-charged bitter substance measurement. Four kinds of modifiers were tested as membrane-modifying materials. By comparing the results obtained from these modifiers, the modifier structure suitable for measuring umami substances was identified. The findings revealed that the presence of carboxyl groups at para-position of the benzene ring, as well as intramolecular H-bonds between the carboxyl group and hydroxyl group, significantly affect the effectiveness of a modifier in the umami substance measurement. The taste sensor treated with this type of modifier showed excellent selectivity for umami substances.
]]>Biosensors doi: 10.3390/bios14020094
Authors: José Alfonso Cruz-Ramos Mijaíl Irak Trapero-Corona Ingrid Aurora Valencia-Hernández Luz Amparo Gómez-Vargas María Teresa Toranzo-Delgado Karla Raquel Cano-Magaña Emmanuel De la Mora-Jiménez Gabriela del Carmen López-Armas
Breast cancer (BC) affects millions of women worldwide, causing over 500,000 deaths annually. It is the leading cause of cancer mortality in women, with 70% of deaths occurring in developing countries. Elastography, which evaluates tissue stiffness, is a promising real-time minimally invasive technique for BC diagnosis. This study assessed strain elastography (SE) and the fat-to-lesion (F/L) index for BC diagnosis. This prospective study included 216 women who underwent SE, ultrasound, mammography, and breast biopsy (108 malignant, 108 benign). Three expert radiologists performed imaging and biopsies. Mean F/L index was 3.70 ± 2.57 for benign biopsies and 18.10 ± 17.01 for malignant. We developed two predictive models: a logistic regression model with AUC 0.893, 79.63% sensitivity, 87.62% specificity, 86.9% positive predictive value (+PV), and 80.7% negative predictive value (−PV); and a neural network with AUC 0.902, 80.56% sensitivity, 88.57% specificity, 87.9% +PV, and 81.6% −PV. The optimal Youden F/L index cutoff was >5.76, with 84.26% sensitivity and specificity. The F/L index positively correlated with BI-RADS (Spearman’s r = 0.073, p < 0.001) and differed among molecular subtypes (Kruskal-Wallis, p = 0.002). SE complements mammography for BC diagnosis. With adequate predictive capacity, SE is fast, minimally invasive, and useful when mammography is contraindicated.
]]>Biosensors doi: 10.3390/bios14020093
Authors: Alexey Ivanov Rezeda Shamagsumova Marina Larina Gennady Evtugyn
Neurodegenerative diseases and Alzheimer’s disease (AD), as one of the most common causes of dementia, result in progressive losses of cholinergic neurons and a reduction in the presynaptic markers of the cholinergic system. These consequences can be compensated by the inhibition of acetylcholinesterase (AChE) followed by a decrease in the rate of acetylcholine hydrolysis. For this reason, anticholinesterase drugs with reversible inhibition effects are applied for the administration of neurodegenerative diseases. Their overdosage, variation in efficiency and recommendation of an individual daily dose require simple and reliable measurement devices capable of the assessment of the drug concentration in biological fluids and medications. In this review, the performance of electrochemical biosensors utilizing immobilized cholinesterases is considered to show their advantages and drawbacks in the determination of anticholinesterase drugs. In addition, common drugs applied in treating neurodegenerative diseases are briefly characterized. The immobilization of enzymes, nature of the signal recorded and its dependence on the transducer modification are considered and the analytical characteristics of appropriate biosensors are summarized for donepezil, huperzine A, rivastigmine, eserine and galantamine as common anti-dementia drugs. Finally, the prospects for the application of AChE-based biosensors in clinical practice are discussed.
]]>Biosensors doi: 10.3390/bios14020092
Authors: Sungcheol Hong Gerard Coté
Pulse Wave Velocity (PWV) analysis is valuable for assessing arterial stiffness and cardiovascular health and potentially for estimating blood pressure cufflessly. However, conventional PWV analysis from two transducers spaced closely poses challenges in data management, battery life, and developing the device for continuous real-time applications together along an artery, which typically need data to be recorded at high sampling rates. Specifically, although a pulse signal consists of low-frequency components when used for applications such as determining heart rate, the pulse transit time for transducers near each other along an artery takes place in the millisecond range, typically needing a high sampling rate. To overcome this issue, in this study, we present a novel approach that leverages the Nyquist–Shannon sampling theorem and reconstruction techniques for signals produced by bioimpedance transducers closely spaced along a radial artery. Specifically, we recorded bioimpedance artery pulse signals at a low sampling rate, reducing the data size and subsequently algorithmically reconstructing these signals at a higher sampling rate. We were able to retain vital transit time information and achieved enhanced precision that is comparable to the traditional high-rate sampling method. Our research demonstrates the viability of the algorithmic method for enabling PWV analysis from low-sampling-rate data, overcoming the constraints of conventional approaches. This technique has the potential to contribute to the development of cardiovascular health monitoring and diagnosis using closely spaced wearable devices for real-time and low-resource PWV assessment, enhancing patient care and cardiovascular disease management.
]]>Biosensors doi: 10.3390/bios14020091
Authors: Hyo Eun Kim Ariadna Schuck Hyeonseek Park Doo Ryeon Chung Minhee Kang Yong-Sang Kim
Despite a substantial increase in testing facilities during the pandemic, access remains a major obstacle, particularly in low-resource and remote areas. This constraint emphasizes the need for high-throughput potential point-of-care diagnostic tools in environments with limited resources. Loop-mediated isothermal amplification (LAMP) is a promising technique, but improvements in sensitivity are needed for accurate detection, especially in scenarios where the virus is present in low quantities. To achieve this objective, we present a highly sensitive detection approach of a dual-mode graphene-based field-effect transistor (G-FET) biosensor with LAMP. The G-FET biosensor, which has a transparent graphene microelectrode array on a glass substrate, detects LAMP products in less than 30 min using both observable color changes and Dirac point voltage measurements, even in samples with low viral concentrations. This dual-mode G-FET biosensor emerges as a potential alternative to conventional RT-PCR for severe acute respiratory syndrome-associated coronavirus (SARS-CoV)-2 detection or point-of-care testing, particularly in resource-constrained scenarios such as developing countries. Moreover, its capacity for colorimetric detection with the naked eye enhances its applicability in diverse settings.
]]>Biosensors doi: 10.3390/bios14020090
Authors: Diana Vitazkova Erik Foltan Helena Kosnacova Michal Micjan Martin Donoval Anton Kuzma Martin Kopani Erik Vavrinsky
This article explores the importance of wearable and remote technologies in healthcare. The focus highlights its potential in continuous monitoring, examines the specificity of the issue, and offers a view of proactive healthcare. Our research describes a wide range of device types and scientific methodologies, starting from traditional chest belts to their modern alternatives and cutting-edge bioamplifiers that distinguish breathing from chest impedance variations. We also investigated innovative technologies such as the monitoring of thorax micromovements based on the principles of seismocardiography, ballistocardiography, remote camera recordings, deployment of integrated optical fibers, or extraction of respiration from cardiovascular variables. Our review is extended to include acoustic methods and breath and blood gas analysis, providing a comprehensive overview of different approaches to respiratory monitoring. The topic of monitoring respiration with wearable and remote electronics is currently the center of attention of researchers, which is also reflected by the growing number of publications. In our manuscript, we offer an overview of the most interesting ones.
]]>Biosensors doi: 10.3390/bios14020089
Authors: Rafael Iriya Brandyn Braswell Manni Mo Fenni Zhang Shelley E. Haydel Shaopeng Wang
Bacterial infections, increasingly resistant to common antibiotics, pose a global health challenge. Traditional diagnostics often depend on slow cell culturing, leading to empirical treatments that accelerate antibiotic resistance. We present a novel large-volume microscopy (LVM) system for rapid, point-of-care bacterial detection. This system, using low magnification (1–2×), visualizes sufficient sample volumes, eliminating the need for culture-based enrichment. Employing deep neural networks, our model demonstrates superior accuracy in detecting uropathogenic Escherichia coli compared to traditional machine learning methods. Future endeavors will focus on enriching our datasets with mixed samples and a broader spectrum of uropathogens, aiming to extend the applicability of our model to clinical samples.
]]>Biosensors doi: 10.3390/bios14020088
Authors: Guanwei Tao Jiajun Li Yunyun Mu Xinping Zhang
We report a three-dimensional (3D) SERS substrate consisting of a silver nanoparticle (AgNP) coating on the skeleton-fiber surfaces of a polytetrafluoroethylene (PTFE) membrane. Simple thermal evaporation was employed to deposit Ag onto the PTFE membrane to produce grape-shaped AgNPs. The 3D-distributed AgNPs exhibit not only strong localized surface plasmon resonance (LSPR) but also strong hydrophobic performance. High-density hotspots via silver nano-grape structures and nanogaps, the large 3D interaction volume, and the large total surface area, in combination with the hydrophobic enrichment of the specimen, facilitate high-sensitivity sensing performance of such a SERS substrate for the direct detection of low-concentration molecules in water. An enhancement factor of up to 1.97 × 1010 was achieved in the direct detection of R6G molecules in water with a concentration of 10−13 mol/L. The lowest detection limit of 100 ppt was reached in the detection of melamine in water. Such a SERS sensor may have potential applications in food-safety control, environmental water pollution monitoring, and biomedical analysis.
]]>Biosensors doi: 10.3390/bios14020087
Authors: Amir Hatamie
“Micro-Nanofabricated Sensors for Bioanalysis” represents a cutting-edge field in biosensing technology which leverages the integration of micro- and nanoscale fabrication techniques [...]
]]>Biosensors doi: 10.3390/bios14020086
Authors: Rohollah Nasiri Yangzhi Zhu Natan Roberto de Barros
The convergence of microfluidics and organ-on-a-chip (OoC) technologies has revolutionized our ability to create advanced in vitro models that recapitulate complex physiological processes [...]
]]>Biosensors doi: 10.3390/bios14020085
Authors: Marion Ryan C. Sytu Andrew Stoner Jong-In Hahm
Mechanical strain has been shown to be a versatile and tunable means to control various properties of nanomaterials. In this work, we investigate how strain applied to individual ZnO nanorods (NRs) can affect the fluorescence signals originated from external sources of bioanalytes, which are subsequently coupled and guided onto the NRs. Specifically, we determine how factors such as the NR length and protein concentration can influence the strain-induced changes in the waveguided fluorescence intensity along the NRs. We employ a protein of tumor necrosis factor-α (TNF-α) and a fluorophore-labeled antibody in a model immunoassay reaction, after which Alexa488-TNF-α immunocomplex is formed on ZnO NRs. We elucidate the relationships between the types as well as amounts of strain on the NRs and the fluorescence intensity originated from the Alexa488-TNF-α immunocomplexes. We show that tensile (compressive) strain applied to the NR leads to an increase (decrease) in the waveguided fluorescence signals. By assessing important optical phenomena such as fluorescence intensification on nanorod ends (FINE) and degree of FINE (DoF), we confirm their linear dependence with both the types and amounts of strain. Furthermore, the strain-induced changes in both FINE and DoF are found to be independent of protein concentration. We determine that NR length plays a critical role in obtaining high strain-dependence of the measured fluorescence signals. Particularly, we ascertain that longer NRs yield larger changes in both FINE and DoF in response to the applied strain, relative to shorter ones. In addition, longer NRs permit higher linear correlation between the protein concentration and the waveguided fluorescence intensity. These outcomes provide valuable insight into exploiting strain to enhance the detection of optical signals from bioanalytes, thus enabling their quantifications even at ultra-trace levels. Coupled with the use of individual ZnO NRs demonstrated in our measurements, this work may contribute to the development of a miniaturized, highly sensitive biosensor whose signal transduction is best optimized by the application of strain.
]]>Biosensors doi: 10.3390/bios14020084
Authors: Jiying Xu Pengfei Zhang Yi Chen
Surface plasmon resonance (SPR) is a powerful tool for determining molecular interactions quantitatively. SPR imaging (SPRi) further improves the throughput of SPR technology and provides the spatially resolved capability for observing the molecular interaction dynamics in detail. SPRi is becoming more and more popular in biological and chemical sensing and imaging. However, SPRi suffers from low spatial resolution due to the imperfect optical components and delocalized features of propagating surface plasmonic waves along the surface. Diverse kinds of approaches have been developed to improve the spatial resolution of SPRi, which have enormously impelled the development of the methodology and further extended its possible applications. In this minireview, we introduce the mechanisms for building a high-spatial-resolution SPRi system and present its experimental schemes from prism-coupled SPRi and SPR microscopy (SPRM) to surface plasmonic scattering microscopy (SPSM); summarize its exciting applications, including molecular interaction analysis, molecular imaging and profiling, tracking of single entities, and analysis of single cells; and discuss its challenges in recent decade as well as the promising future.
]]>Biosensors doi: 10.3390/bios14020083
Authors: Ran An Alireza Avanaki Priyaleela Thota Sai Nemade Amrish Mehta Umut A. Gurkan
Hemoglobin (Hb) disorders are among the most common monogenic diseases affecting nearly 7% of the world population. Among various Hb disorders, approximately 1.5% of the world population carries β-thalassemia (β-Thal), affecting 40,000 newborns every year. Early screening and a timely diagnosis are essential for β-thalassemia patients for the prevention and management of later clinical complications. However, in Africa, Southern Europe, the Middle East, and Southeast Asia, where β-thalassemia is most prevalent, the diagnosis and screening for β-thalassemia are still challenging due to the cost and logistical burden of laboratory diagnostic tests. Here, we present Gazelle, which is a paper-based microchip electrophoresis platform that enables the first point-of-care diagnostic test for β-thalassemia. We evaluated the accuracy of Gazelle for the β-Thal screening across 372 subjects in the age range of 4–63 years at Apple Diagnostics lab in Mumbai, India. Additionally, 30 blood samples were prepared to mimic β-Thal intermediate and β-Thal major samples. Gazelle-detected levels of Hb A, Hb F, and Hb A2 demonstrated high levels of correlation with the results reported through laboratory gold standard high-performance liquid chromatography (HPLC), yielding a Pearson correlation coefficient = 0.99. This ability to obtain rapid and accurate results suggests that Gazelle may be suitable for the large-scale screening and diagnosis of β-Thal.
]]>Biosensors doi: 10.3390/bios14020082
Authors: Isidora P. Gkini Panagiotis Christopoulos Alexis Conides Despina P. Kalogianni Theodore K. Christopoulos
Tuna is an excellent food product, relatively low in calories, that is recommended for a balanced diet. The continuously increasing demand, especially for bluefin-tuna-based food preparations, and its relatively high market price make adulteration by intentionally mixing with other lower-priced tunas more prospective. The development of rapid methods to detect tuna adulteration is a great challenge in food analytical science. We have thus developed a simple, fast, and low-cost molecular rapid test for the visual detection of tuna adulteration. It is the first sensor developed for tuna authenticity testing. The three species studied were Thunnus thynnus (BFT), Thunnus albacares, and Katsuwonus pelamis. DNA was isolated from fresh and heat-treated cooked fish samples followed by PCR. The PCR products were hybridized (10 min) to specific probes and applied to the rapid sensing device. The signal was observed visually in 10–15 min using gold nanoparticle reporters. The method was evaluated employing binary mixtures of PCR products from fresh tissues and mixtures of DNA isolates from heat-treated tissues (canned products) at adulteration percentages of 1–100%. The results showed that the method was reproducible and specific for each tuna species. As low as 1% of tuna adulteration was detected with the naked eye.
]]>Biosensors doi: 10.3390/bios14020081
Authors: Dayle Kotturi Sureyya Paterson Mike McShane
One aim of personalized medicine is to use continuous or on-demand monitoring of metabolites to adjust prescription dosages in real time. Surface-enhanced spatially offset Raman spectroscopy (SESORS) is an optical technique capable of detecting surface-enhanced Raman spectroscopy (SERS)-active targets under a barrier, which may enable frequent metabolite monitoring. Here we investigate how the intensity of the signal from SERS-active material varies spatially through tissue, both experimentally and in a computational model. Implant-sized, SERS-active hydrogel was placed under different thicknesses of contiguous tissue. Emission spectra were collected at the air-tissue boundary over a range of offsets from the excitation site. New features were added to the Monte Carlo light-tissue interaction model to modify the optical properties after inelastic scattering and to calculate the distribution of photons as they exit the model. The Raman signals were detectable through all barrier thicknesses, with strongest emission for the case of 0 mm offset between the excitation and detector. A steep decline in the signal intensities occurred for offsets greater than 2 mm. These results did not match published SORS work (where targets were much larger than an implant). However, the model and experimental results agree in showing the greatest intensities at 0 mm offset and a steep gradient in the intensities with increasing offset. Also, the model showed an increase in the number of photons when the new, longer wavelengths were used following the Stokes shift for scattering and the graphical display of the exiting photons was helpful in the determination and confirmation of the optimal offset.
]]>Biosensors doi: 10.3390/bios14020080
Authors: Andy Chieng Zijian Wan Shaopeng Wang
The detection and analysis of small molecules, typically defined as molecules under 1000 Da, is of growing interest ranging from the development of small-molecule drugs and inhibitors to the sensing of toxins and biomarkers. However, due to challenges such as their small size and low mass, many biosensing technologies struggle to have the sensitivity and selectivity for the detection of small molecules. Notably, their small size limits the usage of labeled techniques that can change the properties of small-molecule analytes. Furthermore, the capability of real-time detection is highly desired for small-molecule biosensors’ application in diagnostics or screening. This review highlights recent advances in label-free real-time biosensing technologies utilizing different types of transducers to meet the growing demand for small-molecule detection.
]]>Biosensors doi: 10.3390/bios14020079
Authors: Francesco Lavecchia di Tocco Valentina Botti Salvatore Cannistraro Anna Rita Bizzarri
MicroRNAs are small ribonucleotides that act as key gene regulators. Their altered expression is often associated with the onset and progression of several human diseases, including cancer. Given their potential use as biomarkers, there is a need to find detection methods for microRNAs suitable for use in clinical setting. Field-effect-transistor-based biosensors (bioFETs) appear to be valid tools to detect microRNAs, since they may reliably quantitate the specific binding between the immobilized probe and free target in solution through an easily detectable electrical signal. We have investigated the detection of human microRNA 155 (miR-155) using an innovative capturing probe constituted by a synthetic peptide nucleic acid (PNA), which has the advantage to form a duplex even at ionic strengths approaching the physiological conditions. With the aim to develop an optimized BioFET setup, the interaction kinetics between miR-155 and the chosen PNA was preliminarily investigated by using surface plasmon resonance (SPR). By exploiting both these results and our custom-made bioFET system, we were able to attain a low-cost, real-time, label-free and highly specific detection of miR-155 in the nano-molar range.
]]>Biosensors doi: 10.3390/bios14020078
Authors: Arash Mohammadinejad Laura Elena Gaman Ghazaleh Aleyaghoob Liviu Gaceu Seyed Ahmad Mohajeri Marius Alexandru Moga Mihaela Badea
Cancer is known as one of the most significant causes of death worldwide, and, in spite of novel therapeutic methods, continues to cause a considerable number of deaths. Targeted molecular diagnosis and therapy using aptamers with high affinity have become popular techniques for pathological angiogenesis and cancer therapy scientists. In this paper, several aptamer-based diagnostic and therapeutic techniques such as aptamer–nanomaterial conjugation, aptamer–drug conjugation (physically or covalently), and biosensors, which have been successfully designed for biomarkers, were critically reviewed. The results demonstrated that aptamers can potentially be incorporated with targeted delivery systems and biosensors for the detection of biomarkers expressed by cancer cells. Aptamer-based therapeutic and diagnostic methods, representing the main field of medical sciences, possess high potential for use in cancer therapy, pathological angiogenesis, and improvement of community health. The clinical use of aptamers is limited due to target impurities, inaccuracy in the systematic evolution of ligands via exponential enrichment (SELEX)stage process, and in vitro synthesis, making them unreliable and leading to lower selectivity for in vivo targets. Moreover, size, behavior, probable toxicity, low distribution, and the unpredictable behavior of nanomaterials in in vivo media make their usage in clinical assays critical. This review is helpful for the implementation of aptamer-based therapies which are effective and applicable for clinical use and the design of future studies.
]]>Biosensors doi: 10.3390/bios14020077
Authors: Noha Elnagar Nada Elgiddawy Waleed M. A. El Rouby Ahmed A. Farghali Hafsa Korri-Youssoufi
The sensitive determination of folate receptors (FRs) in the early stages of cancer is of great significance for controlling the progression of cancerous cells. Many folic acid (FA)-based electrochemical biosensors have been utilized to detect FRs with promising performances, but most were complicated, non-reproducible, non-biocompatible, and time and cost consuming. Here, we developed an environmentally friendly and sensitive biosensor for FR detection. We proposed an electrochemical impedimetric biosensor formed by nanofibers (NFs) of bio-copolymers prepared by electrospinning. The biosensor combines the advantages of bio-friendly polymers, such as sodium alginate (SA) and polyethylene oxide (PEO) as an antifouling polymer, with FA as a biorecognition element. The NF nanocomposites were characterized using various techniques, including SEM, FTIR, zeta potential (ZP), cyclic voltammetry (CV), and electrochemical impedance spectroscopy (EIS). We evaluated the performance of the NF biosensor using EIS and demonstrated FR detection in plasma with a limit of detection of 3 pM. Furthermore, the biosensor showed high selectivity, reliability, and good stability when stored for two months. This biosensor was constructed from ‘green credentials’ holding polymers that are highly needed in the new paradigm shift in the medical industry.
]]>Biosensors doi: 10.3390/bios14020076
Authors: Vladislava Kapravchuk Andrey Briko Alexander Kobelev Ahmad Hammoud Sergey Shchukin
This present work is aimed at conducting fundamental and exploratory studies of the mechanisms of electrical impedance signal formation. This paper also considers morphofunctional changes in forearm tissues during the performance of basic hand actions. For this purpose, the existing research benches were modernized to conduct experiments of mapping forearm muscle activity by electrode systems on the basis of complexing the electrical impedance signals and electromyography signals and recording electrode systems’ pressing force using force transducers. Studies were carried out with the involvement of healthy volunteers in the implementation of vertical movement of the electrode system and ultrasound transducer when the subject’s upper limb was positioned in the bed of the stand while performing basic hand actions in order to identify the relationship between the morphofunctional activity of the upper limb muscles and the recorded parameters of the electro-impedance myography signal. On the basis of the results of the studies, including complex measurements of neuromuscular activity on healthy volunteers such as the signals of electro-impedance myography and pressing force, analyses of the morphofunctional changes in tissues during action performance on the basis of ultrasound and MRI studies and the factors influencing the recorded signals of electro-impedance myography are described. The results are of fundamental importance and will enable reproducible electro-impedance myography signals, which, in turn, allow improved anthropomorphic control.
]]>Biosensors doi: 10.3390/bios14020075
Authors: Rosalía González Brito Pablo Montenegro Alicia Méndez Ramtin E. Shabgahi Alberto Pasquarelli Ricardo Borges
Amperometry is arguably the most widely used technique for studying the exocytosis of biological amines. However, the scarcity of human tissues, particularly in the context of neurological diseases, poses a challenge for exocytosis research. Human platelets, which accumulate 90% of blood serotonin, release it through exocytosis. Nevertheless, single-cell amperometry with encapsulated carbon fibers is impractical due to the small size of platelets and the limited number of secretory granules on each platelet. The recent technological improvements in amperometric multi-electrode array (MEA) devices allow simultaneous recordings from several high-performance electrodes. In this paper, we present a comparison of three MEA boron-doped diamond (BDD) devices for studying serotonin exocytosis in human platelets: (i) the BDD-on-glass MEA, (ii) the BDD-on-silicon MEA, and (iii) the BDD on amorphous quartz MEA (BDD-on-quartz MEA). Transparent electrodes offer several advantages for observing living cells, and in the case of platelets, they control activation/aggregation. BDD-on-quartz offers the advantage over previous materials of combining excellent electrochemical properties with transparency for microscopic observation. These devices are opening exciting perspectives for clinical applications.
]]>Biosensors doi: 10.3390/bios14020074
Authors: Benjamin C. Kreager Huaiyu Wu Wei-Yi Chang Sunho Moon Josh Mitchell Chang Peng Chih-Chung Huang Marie Muller Jian Tian Xiaoning Jiang
To address the need for high-resolution imaging in lung nodule detection and overcome the limitations of the shallow imaging depth associated with high-frequency ultrasound and the complex structure of lung tissue, we successfully integrated 50 MHz ultrasound transducers with 18-gauge biopsy needles. Featuring a miniaturized size of 0.6 × 0.5 × 0.5 mm3, the 50 MHz micromachined 1-3 composite transducer was tested to perform mechanical scanning of a nodule within a lung-tissue-mimicking phantom in vitro. The high-frequency transducer demonstrated the ability to achieve imaging with an axial resolution of 30 μm for measuring nodule edges. Moreover, the integrated biopsy needle prototype exhibited high accuracy (1.74% discrepancy) in estimating nodule area compared to actual dimensions in vitro. These results underscore the promising potential of biopsy-needle-integrated transducers in enhancing the accuracy of endoscopic ultrasound-guided fine needle aspiration biopsy (EUS-FNA) for clinical applications.
]]>Biosensors doi: 10.3390/bios14020073
Authors: Da Eun Oh Hyun Beom Kim Tae Hyun Kim
An electrochemically reduced graphene oxide (ERGO) electrode-based electrochemical assay was developed for rapid, sensitive, and straightforward analysis of both activity and inhibition of the endonuclease EcoRV. The procedure uses a DNA substrate designed for EcoRV, featuring a double-stranded DNA (dsDNA) region labeled with methylene blue (MB) and a single-stranded DNA (ssDNA) region immobilized on the ERGO surface. The ERGO electrode, immobilized with the DNA substrate, was subsequently exposed to a sample containing EcoRV. Upon enzymatic hydrolysis, the cleaved dsDNA fragments were detached from the ERGO surface, leading to a decrease in the MB concentration near the electrode. This diminished the electron transfer efficiency for MB reduction, resulting in a decreased reduction current. This assay demonstrates excellent specificity and high sensitivity, with a limit of detection (LOD) of 9.5 × 10−3 U mL−1. Importantly, it can also measure EcoRV activity in the presence of aurintricarboxylic acid, a known inhibitor, highlighting its potential for drug discovery and clinical diagnostic applications.
]]>Biosensors doi: 10.3390/bios14020072
Authors: Lorenzo Nardi Nithin Maipan Davis Serena Sansolini Thiago Baratto de Albuquerque Mohcine Laarraj Domenico Caputo Giampiero de Cesare Seyedeh Rojin Shariati Pour Martina Zangheri Donato Calabria Massimo Guardigli Michele Balsamo Elisa Carrubba Fabrizio Carubia Marco Ceccarelli Michele Ghiozzi Liyana Popova Andrea Tenaglia Marino Crisconio Alessandro Donati Augusto Nascetti Mara Mirasoli
One of the main challenges to be faced in deep space missions is to protect the health and ensure the maximum efficiency of the crew by preparing methods of prevention and in situ diagnosis. Indeed, the hostile environment causes important health problems, ranging from muscle atrophy, osteopenia, and immunological and metabolic alterations due to microgravity, to an increased risk of cancer caused by exposure to radiation. It is, therefore, necessary to provide new methods for the real-time measurement of biomarkers suitable for deepening our knowledge of the effects of space flight on the balance of the immune system and for allowing the monitoring of the astronaut’s health during long-term missions. APHRODITE will enable human space exploration because it fills this void that affects both missions in LEO and future missions to the Moon and Mars. Its scientific objectives are the design, production, testing, and in-orbit demonstration of a compact, reusable, and reconfigurable system for performing the real-time analysis of oral fluid samples in manned space missions. In the frame of this project, a crew member onboard the ISS will employ APHRODITE to measure the selected target analytes, cortisol, and dehydroepiandrosterone sulfate (DHEA-S), in oral fluid, in four (plus one additional desired session) separate experiment sessions. The paper addresses the design of the main subsystems of the analytical device and the preliminary results obtained during the first implementations of the device subsystems and testing measurements on Earth. In particular, the system design and the experiment data output of the lab-on-chip photosensors and of the front-end readout electronics are reported in detail along with preliminary chemical tests for the duplex competitive CL-immunoassay for the simultaneous detection of cortisol and DHEA-S. Different applications also on Earth are envisaged for the APHRODITE device, as it will be suitable for point-of-care testing applications (e.g., emergency medicine, bioterrorism, diagnostics in developing countries, etc.).
]]>Biosensors doi: 10.3390/bios14020071
Authors: Akinrinade George Ayankojo Jekaterina Reut Vitali Syritski
Early-stage detection and diagnosis of diseases is essential to the prompt commencement of treatment regimens, curbing the spread of the disease, and improving human health. Thus, the accurate detection of disease biomarkers through the development of robust, sensitive, and selective diagnostic tools has remained cutting-edge scientific research for decades. Due to their merits of being selective, stable, simple, and having a low preparation cost, molecularly imprinted polymers (MIPs) are increasingly becoming artificial substitutes for natural receptors in the design of state-of-the-art sensing devices. While there are different MIP preparation approaches, electrochemical synthesis presents a unique and outstanding method for chemical sensing applications, allowing the direct formation of the polymer on the transducer as well as simplicity in tuning the film properties, thus accelerating the trend in the design of commercial MIP-based sensors. This review evaluates recent achievements in the applications of electrosynthesized MIP sensors for clinical analysis of disease biomarkers, identifying major trends and highlighting interesting perspectives on the realization of commercial MIP-endowed testing devices for rapid determination of prevailing diseases.
]]>Biosensors doi: 10.3390/bios14020070
Authors: Vince S. Siu Minhua Lu Kuan Yu Hsieh Bo Wen Italo Buleje Nigel Hinds Krishna Patel Bing Dang Russell Budd
This paper presents a cost-effective, quantitative, point-of-care solution for urinalysis screening, specifically targeting nitrite, protein, creatinine, and pH in urine samples. Detecting nitrite is crucial for the early identification of urinary tract infections (UTIs), while regularly measuring urinary protein-to-creatinine (UPC) ratios aids in managing kidney health. To address these needs, we developed a portable, transmission-based colorimeter using readily available components, controllable via a smartphone application through Bluetooth. Multiple colorimetric detection strategies for each analyte were identified and tested for sensitivity, specificity, and stability in a salt buffer, artificial urine, and human urine. The colorimeter successfully detected all analytes within their clinically relevant ranges: nitrite (6.25–200 µM), protein (2–1024 mg/dL), creatinine (2–1024 mg/dL), and pH (5.0–8.0). The introduction of quantitative protein and creatinine detection, and a calculated urinary protein-to-creatinine (UPC) ratio at the point-of-care, represents a significant advancement, allowing patients with proteinuria to monitor their condition without frequent lab visits. Furthermore, the colorimeter provides versatile data storage options, facilitating local storage on mobile devices or in the cloud. The paper further details the setup of the colorimeter’s secure connection to a cloud-based environment, and the visualization of time-series analyte measurements in a web-based dashboard.
]]>Biosensors doi: 10.3390/bios14020069
Authors: Zhichao Li Hongbin Pu Qingyi Wei
Dimethoate contaminants in food pose a threat to human health. Rapid and sensitive trace detection methods are required to keep food safe. In this study, a novel fluorescent aptasensor was developed for the sensitive detection of dimethoate based on carbon quantum dots labeled with double-stranded DNA (CQDs−apt−cDNA) and Ti3C2Tx flakes. Under optimal conditions, the aptasensor showed a good linear range of 1 × 10−9 to 5 × 10−5 M for dimethoate with a coefficient of determination (R2) of 0.996. Besides, a low detection limit of 2.18 × 10−10 M was obtained. The aptasensor showed high selectivity in interference samples and good reproducibility with an RSD of 3.06% (<5%) for dimethoate detection. Furthermore, the proposed aptasensor was applied to the detection of dimethoate in apple juice and tap water with satisfactory recoveries from 96.2 to 104.4%. Because of these benefits, this aptasensor has the potential and promise for detecting food contaminants in the food industry.
]]>Biosensors doi: 10.3390/bios14020068
Authors: Grace M. Hwang Aleksandr L. Simonian
In partnership with the Air Force Office of Scientific Research (AFOSR), the National Science Foundation’s (NSF) Emerging Frontiers and Multidisciplinary Activities (EFMA) office of the Directorate for Engineering (ENG) launched an Emerging Frontiers in Research and Innovation (EFRI) topic for the fiscal years FY22 and FY23 entitled “Brain-inspired Dynamics for Engineering Energy-Efficient Circuits and Artificial Intelligence” (BRAID) [...]
]]>Biosensors doi: 10.3390/bios14020067
Authors: Norica-Carmen Godja Florentina-Daniela Munteanu
The integration of nanomaterials into sensor technologies not only poses challenges but also opens up promising prospects for future research. These challenges include assessing the toxicity of nanomaterials, scalability issues, and the seamless integration of these materials into existing infrastructures. Future development opportunities lie in creating multifunctional nanocomposites and environmentally friendly nanomaterials. Crucial to this process is collaboration between universities, industry, and regulatory authorities to establish standardization in this evolving field. Our perspective favours using screen-printed sensors that employ nanocomposites with high electrochemical conductivity. This approach not only offers cost-effective production methods but also allows for customizable designs. Furthermore, incorporating hybrids based on carbon-based nanomaterials and functionalized Mxene significantly enhances sensor performance. These high electrochemical conductivity sensors are portable, rapid, and well-suited for on-site environmental monitoring, seamlessly aligning with Internet of Things (IoT) platforms for developing intelligent systems. Simultaneously, advances in electrochemical sensor technology are actively working to elevate sensitivity through integrating nanotechnology, miniaturization, and innovative electrode designs. This comprehensive approach aims to unlock the full potential of sensor technologies, catering to diverse applications ranging from healthcare to environmental monitoring. This review aims to summarise the latest trends in using hybrid nanomaterial-based sensors, explicitly focusing on their application in detecting environmental contaminants.
]]>Biosensors doi: 10.3390/bios14020066
Authors: Donatas Pelenis Dovydas Barauskas Mindaugas Dzikaras Darius Viržonis
Custom electronics tailored for ultrasonic applications with four ultrasonic transmit-receive channels and a nominal 25 MHz single channel frequency were developed for ultrasound BAW and SAW biosensor uses. The designed integrated microcontroller, supported by Python with a SciPy library, and the developed system measured the time of flight (TOF) and other wave properties to characterize the acoustic properties of a bulk of the liquid in a microchannel or acoustic properties of biological species attached to an analytic surface in real time. The system can utilize both piezoelectric and capacitive micromachined ultrasound transducers. The device demonstrated a linear response to changes in water salinity. This response was primarily attributed to the time-of-flight (TOF) changes related to the varying solution density. Furthermore, real-time DNA oligonucleotide-based interactions between oligonucleotides immobilized on the device’s analytical area and oligonucleotides attached to gold nanoparticles (Au NPs) in the solution were demonstrated. The biological interaction led to an exponential decrease in the acoustic interfacial wave propagating across the interface between the solution and the solid surface of the sensor, the TOF signal. This decrease was attributed to the increase in the effective density of the solution in the vicinity of the sensor’s analytical area, as Au NPs modified by oligonucleotides were binding to the analytical area. The utilization of Au NPs in oligonucleotide surface binding yields a considerably stronger sensor signal than previously observed in earlier CMUT-based TOF biosensor prototypes.
]]>Biosensors doi: 10.3390/bios14020065
Authors: Jung-Chih Lin Zhao-Yu Zhou Yi-Ching Cheng I-Nan Chang Chu-En Lin Chi-Chang Wu
Field-effect transistor (FET)-based biosensors are powerful analytical tools for detecting trace-specific biomolecules in diverse sample matrices, especially in the realms of pandemics and infectious diseases. The primary concern in applying these biosensors is their stability, a factor directly impacting the accuracy and reliability of sensing over extended durations. The risk of biosensor degradation is substantial, potentially jeopardizing the sensitivity and selectivity and leading to inaccurate readings, including the possibility of false positives or negatives. This paper delves into the documented degradation of silicon nanobelt FET (NBFET) biosensors induced by buffer solutions. The results highlight a positive correlation between immersion time and the threshold voltage of NBFET devices. Secondary ion mass spectrometry analysis demonstrates a gradual increase in sodium and potassium ion concentrations within the silicon as immersion days progress. This outcome is ascribed to the nanobelt’s exposure to the buffer solution during the biosensing period, enabling ion penetration from the buffer into the silicon. This study emphasizes the critical need to address buffer-solution-induced degradation to ensure the long-term stability and performance of FET-based biosensors in practical applications.
]]>Biosensors doi: 10.3390/bios14020063
Authors: Yang Guo Jingran Sun Mingzhu Liu Jin Wu Zunquan Zhao Ting Ma Yanjun Fang
Nanomaterials are desirable for sensing applications. Therefore, MnO2 nanosheets and nitrogen-doped carbon dots (NCDs) were used to construct a ratiometric biosensor for quantification of 2,4-dichlorophenoxyacetic acid. The MnO2 nanosheets drove the oxidation of colorless o-phenylenediamine to OPDox, which exhibits fluorescence emission peaks at 556 nm. The fluorescence of OPDox was efficiently quenched and the NCDs were recovered as the ascorbic acid produced by the hydrolyzed alkaline phosphatase (ALP) substrate increased. Owing to the selective inhibition of ALP activity by 2,4-D and the inner filter effect, the fluorescence intensity of the NCDs at 430 nm was suppressed, whereas that at 556 nm was maintained. The fluorescence intensity ratio was used for quantitative detection. The linear equation was F = 0.138 + 3.863·C 2,4-D (correlation coefficient R2 = 0.9904), whereas the limits of detection (LOD) and quantification (LOQ) were 0.013 and 0.040 μg/mL. The method was successfully employed for the determination of 2,4-D in different vegetables with recoveries of 79%~105%. The fluorescent color change in the 2,4-D sensing system can also be captured by a smartphone to achieve colorimetric detection by homemade portable test kit.
]]>Biosensors doi: 10.3390/bios14020064
Authors: Weibo Fang Xudong Liu Mariam Maiga Wenjian Cao Ying Mu Qiang Yan Qiangyuan Zhu
Single-cell analysis provides an overwhelming strategy for revealing cellular heterogeneity and new perspectives for understanding the biological function and disease mechanism. Moreover, it promotes the basic and clinical research in many fields at a single-cell resolution. A digital polymerase chain reaction (dPCR) is an absolute quantitative analysis technology with high sensitivity and precision for DNA/RNA or protein. With the development of microfluidic technology, digital PCR has been used to achieve absolute quantification of single-cell gene expression and single-cell proteins. For single-cell specific-gene or -protein detection, digital PCR has shown great advantages. So, this review will introduce the significance and process of single-cell analysis, including single-cell isolation, single-cell lysis, and single-cell detection methods, mainly focusing on the microfluidic single-cell digital PCR technology and its biological application at a single-cell level. The challenges and opportunities for the development of single-cell digital PCR are also discussed.
]]>Biosensors doi: 10.3390/bios14020062
Authors: Chengzhen Ji Xuemei Tang Ruiming Wen Chengdong Xu Jing Wei Bingjun Han Long Wu
Ethephon (ETH), a commonly employed growth regulator, poses potential health risks due to its residue in fruits and vegetables, leading to both acute and subchronic toxicity. However, the detection accuracy of ETH is compromised by the color effects of the samples during the detection process. In this work, a multienzyme reaction-mediated electrochemical biosensor (MRMEC) was developed for the sensitive, rapid, and color-interference-resistant determination of ETH. Nanozymes Fe3O4@Au–Pt and graphene nanocomplexes (GN–Au NPs) were prepared as catalysts and signal amplifiers for MRMEC. Acetylcholinesterase (AChE), acetylcholine (ACh), and choline oxidase (CHOx) form a cascade enzyme reaction to produce H2O2 in an electrolytic cell. Fe3O4@Au–Pt has excellent peroxidase-like activity and can catalyze the oxidation of 3,3′,5,5′-tetramethvlbenzidine (TMB) in the presence of H2O2, resulting in a decrease in the characteristic peak current of TMB. Based on the inhibitory effect of ETH on AChE, the differential pulse voltammetry (DPV) current signal of TMB was used to detect ETH, offering the limit of detection (LOD) of 2.01 nmol L−1. The MRMEC method effectively analyzed ETH levels in mangoes, showing satisfactory precision (coefficient of variations, 2.88–15.97%) and recovery rate (92.18–110.72%). This biosensor holds promise for detecting various organophosphorus pesticides in food samples.
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