In recent years, the biochemical and biological research areas have shown great interest in a smart wearable sensor because of its increasing prevalence and high potential to monitor human health in a non-invasive manner by continuous screening of biomarkers dispersed throughout the biological analytes, as well as real-time diagnostic tools and time-sensitive information compared to conventional hospital-centered system. These smart wearable sensors offer an innovative option for evaluating and investigating human health by incorporating a portion of recent advances in technology and engineering that can enhance real-time point-of-care-testing capabilities. Smart wearable sensors have emerged progressively with a mixture of multiplexed biosensing, microfluidic sampling, and data acquisition systems incorporated with flexible substrate and bodily attachments for enhanced wearability, portability, and reliability. There is a good chance that smart wearable sensors will be relevant to the early detection and diagnosis of disease management and control. Therefore, pioneering smart wearable sensors into reality seems extremely promising despite possible challenges in this cutting-edge technology for a better future in the healthcare domain. This review presents critical viewpoints on recent developments in wearable sensors in the upcoming smart digital health monitoring in real-time scenarios. In addition, there have been proactive discussions in recent years on materials selection, design optimization, efficient fabrication tools, and data processing units, as well as their continuous monitoring and tracking strategy with system-level integration such as internet-of-things, cyber-physical systems, and machine learning algorithms.
Wearable Electronic Devices , Humans , Reproducibility of Results , Point-of-Care Testing , Digital Health , Technology
This article reviews the recent advances in the field of batteryless near-field communication (NFC) sensors for chemical sensing and biosensing. The commercial availability of low-cost commercial NFC integrated circuits (ICs) and their massive integration in smartphones, used as readers and cloud interfaces, have aroused great interest in new batteryless NFC sensors. The fact that coil antennas are not importantly affected by the body compared with other wireless sensors based on far-field communications makes this technology suitable for future wearable point-of-care testing (PoCT) devices. This review first compares energy harvesting based on NFC to other energy-harvesting technologies. Next, some practical recommendations for designing and tuning NFC-based tags are described. Power transfer is key because in most cases, the energy harvested has to be stable for several seconds and not contaminated by undesired signals. For this reason, the effect of the dimensions of the coils and the conductivity on the wireless power transfer is thoroughly discussed. In the last part of the review, the state of the art in NFC-based chemical and biosensors is presented. NFC-based tags (or sensor tags) are mainly based on commercial or custom NFC ICs, which are used to harvest the energy from the RF field generated by the smartphone to power the electronics. Low-consumption colorimeters and potentiostats can be integrated into these NFC tags, opening the door to the integration of chemical sensors and biosensors, which can be harvested and read from a smartphone. The smartphone is also used to upload the acquired information to the cloud to facilitate the internet of medical things (IoMT) paradigm. Finally, several chipless sensors recently proposed in the literature as a low-cost alternative for chemical applications are discussed.
Communication , Internet of Things , Electric Conductivity , Electronics , Point-of-Care Testing
The development of simple, cost-effective, rapid, and quantitative diagnostic tools remains critical to monitor infectious COVID-19 disease. Although numerous diagnostic platforms, including rapid antigen tests, are developed and used, they suffer from limited accuracy, especially when tested with asymptomatic patients. Here, a unique approach to fabricate a nanochannel-based electrochemical biosensor that can detect the entire virion instead of virus fragments, is demonstrated. The sensing platform has uniform nanoscale channels created by the convective assembly of polystyrene (PS) beads on gold electrodes. The PS beads are then functionalized with bioreceptors while the gold surface is endowed with anti-fouling properties. When added to the biosensor, SARS-CoV-2 virus particles block the nanochannels by specific binding to the bioreceptors. The nanochannel blockage hinders the diffusion of a redox probe; and thus, allows quantification of the viral load by measuring the changes in the oxidation current before and after virus incubation. The biosensor shows a low limit of detection of ≈1.0 viral particle mL-1 with a wide detection range up to 108 particles mL-1 in cell culture media. Moreover, the biosensor is able to differentiate saliva samples with SARS-CoV-2 from those without, demonstrating the potential of this technology for translation into a point-of-care biosensor product.
Mesoporous thin films are widely used for applications in need of high surface area and efficient mass and charge transport properties. A well-established fabrication process involves the supramolecular assembly of organic molecules (e.g., block copolymers and surfactants) with inorganic materials obtained by sol-gel chemistry. Typically, subsequent calcination in air removes the organic template and reveals the porous inorganic network. A significant challenge for such coatings is the anisotropic shrinkage due to the volume contraction related to solvent evaporation, inorganic condensation, and template removal, affecting the final porosity as well as pore shape, size, arrangement, and accessibility. Here, we show that a two-step calcination process, composed of high-temperature treatment in argon followed by air calcination, is an effective fabrication strategy to reduce film contraction and enhance structural control of mesoporous thin films. Crucially, the formation of a transient carbonaceous scaffold enables the inorganic matrix to fully condense before template removal. The resulting mesoporous films retain a higher porosity as well as bigger pores with extended porous order. Such films present favorable characteristics for mass transport of large molecules. This is demonstrated for lysozyme adsorption into the mesoporous thin films as an example of enzyme storage.
The methyl erythritol phosphate (MEP) pathway of isoprenoid biosynthesis is essential for malaria parasites and also for several human pathogenic bacteria, thus representing an interesting target for future antimalarials and antibiotics and for diagnostic strategies. We have developed a DNA aptamer (D10) against Plasmodium falciparum 1-deoxy-D-xylulose-5-phosphate reductoisomerase (DXR), the second enzyme of this metabolic route. D10 binds in vitro to recombinant DXR from P. falciparum and Escherichia coli, showing at 10 µM a ca. 50% inhibition of the bacterial enzyme. In silico docking analysis indicates that D10 associates with DXR in solvent-exposed regions outside the active center pocket. According to fluorescence confocal microscopy data, this aptamer specifically targets in P. falciparum in vitro cultures the apicoplast organelle where the MEP pathway is localized and is, therefore, a highly specific marker of red blood cells parasitized by Plasmodium vs. naïve erythrocytes. D10 is also selective for the detection of MEP+ bacteria (e.g., E. coli and Pseudomonas aeruginosa) vs. those lacking DXR (e.g., Enterococcus faecalis). Based on these results, we discuss the potential of DNA aptamers in the development of ligands that can outcompete the performance of the well-established antibody technology for future therapeutic and diagnostic approaches.
This work studies the feasibility of using a battery-less Near-Field Communication (NFC) potentiostat for the next generation of electrochemical point-of-care sensors. A design based on an NFC microchip, a microcontroller, and a custom potentiostat based on an operational amplifier is presented. A proof-of-concept prototype has been designed and used to quantify glucose concentration using commercial glucose test strips from chronoamperometry measurements. The device is harvested and the sensor is read using a mobile phone. The prototype uses an antenna loop covered with ferrite sheets to ensure stable operation of the electronics when the mobile phone is used as reader. The use of ferrite reduces the detuning caused by the proximity of the metal parts of the mobile phone. A comparison with a commercial glucometer device is provided. Results obtained using a commercial glucometer and those provided by the proposed potentiostat show an excellent agreement.
Electric Power Supplies , Point-of-Care Systems , Ferric Compounds , Glucose
Rapid, sensitive, selective and portable virus detection is in high demand globally. However, differentiating non-infectious viral particles from intact/infectious viruses is still a rarely satisfied sensing requirement. Using the negative space within monolayers of polystyrene (PS) spheres deposited directly on gold electrodes, we fabricated tuneable nanochannels decorated with target-selective bioreceptors that facilitate the size-selective detection of intact viruses. Detection occurred through selective nanochannel blockage of diffusion of a redox probe, [Fe(CN)6]3/4-, allowing a quantifiable change in the oxidation current before and after analyte binding to the bioreceptor immobilised on the spheres. Our model system involved partial surface passivation of the mono-assembled PS spheres, by silica glancing angle deposition, to confine bioreceptor immobilisation specifically to the channels and improve particle detection sensitivity. Virus detection was first optimised and modelled with biotinylated gold nanoparticles, recognised by streptavidin immobilised on the PS layer, reaching a low limit of detection of 37 particles/mL. Intact, label-free virus detection was demonstrated using MS2 bacteriophage (~23-28 nm), a marker of microbiological contamination, showing an excellent limit of detection of ~1.0 pfu/mL. Tuneable nanochannel geometries constructed directly on sensing electrodes offer label-free, sensitive, and cost-efficient point-of-care biosensing platforms that could be applied for a wide range of viruses.
Biosensing Techniques , Metal Nanoparticles , Electrochemical Techniques , Electrodes , Gold/chemistry , Limit of Detection , Metal Nanoparticles/chemistry
In this work, a novel, cost-effective, and eco-friendly electrochemical (EC) nanosensor was fabricated for the simultaneous detection of daptomycin (DAP) and meropenem (MEROP). EC methods have been developed for the determination of antibiotics. In this context, green synthesized copper nanoparticles (CuNPs) using Moringa oleifera plant extract were used as electrode modifiers. The incorporation of CuNPs was proposed to enhance the sensitivity and allow the simultaneous quantification of both antibiotics in water. Transmission electron microscopy (TEM), dynamic light scattering (DLS), attenuated total reflectance Fourier transform infrared (ATR-FTIR) spectroscopy, UV-visible spectroscopy, and field emission scanning electron microscopy with energy dispersive X-ray spectroscopy (FESEM-EDX) were employed to characterize CuNPs. Physical adsorption of 20.0 nm (±2.2 nm) spherical CuNPs on the surface of screen-printed carbon electrodes (SPCEs) induced a remarkable electrocatalytic effect. Indeed, the detection of both antibiotics exhibited a limit of detection (LOD) of 0.01 g L-1. The response to various interfering species was assessed. Finally, the quantification of DAP and MEROP in drinking water was demonstrated, confirming the potential of the developed sensor for environmental monitoring applications.
Drinking Water , Nanoparticles , Anti-Bacterial Agents/pharmacology , Copper/chemistry , Copper/pharmacology , Electrochemical Techniques
In naturally occurring bovine mastitis, effects of infection depend on the host inflammatory response, including the effects of secreted cytokines. Knowledge about the inflammatory and regulatory cytokines in milk cells of free-stall barn dairy cows and in naturally occurring mastitis is lacking as most studies focus on induced mastitis. Hereby, the aim of the study was to determine inflammatory and regulatory cytokines in the milk of dairy cows with subclinical and clinical mastitis. The following examinations of milk samples were performed: differential counting of somatic cells (SCC), bacteriological examination, and immunocytochemical analysis. Mean SCC increased in subclinical and clinical mastitis cases. The number of pathogenic mastitis-causing bacteria on plates increased in subclinical mastitis cases but decreased in clinical mastitis. The inflammatory and regulatory markers in the milk cells of healthy cows showed the highest mean cell numbers (%). In mastitis cases, immunoreactivity was more pronounced for IL-4, IL-6, IL-12, IL-13, IL-17A, TNF-α, and IFN-γ. Data about subclinical and clinical mastitis demonstrate inflammatory responses to intramammary infection driven by IL-1α, IL-4, and IL-17A. Moreover, the host defense response in mastitis is characterized by continuation or resolution of initial inflammation. IL-12 and INF-γ immunoreactivity was recognized to differ mastitis cases from the relative health status.
Porous silicon (pSi) is an established porous material that offers ample opportunities for biosensor design thanks to its tunable structure, versatile surface chemistry, and large surface area. Nonetheless, its potential for electrochemical sensing is relatively unexplored. This study investigates layered carbon-stabilized pSi nanostructures with site-specific functionalities as an electrochemical biosensor. A double-layer nanostructure combining a top hydrophilic layer of thermally carbonized pSi (TCpSi) and a bottom hydrophobic layer of thermally hydrocarbonized pSi (THCpSi) is prepared. The modified layers are formed in a stepwise process, involving first an electrochemical anodization step to generate a porous layer with precisely defined pore morphological features, followed by deposition of a thin thermally carbonized coating on the pore walls via temperature-controlled acetylene decomposition. The second layer is then generated beneath the first by following the same two-step process, but the acetylene decomposition conditions are adjusted to deposit a thermally hydrocarbonized coating. The double-layer platform features excellent electrochemical properties such as fast electron-transfer kinetics, which underpin the performance of a TCpSi-THCpSi voltammetric DNA sensor. The biosensor targets a 28-nucleotide single-stranded DNA sequence with a detection limit of 0.4 pM, two orders of magnitude lower than the values reported to date by any other pSi-based electrochemical DNA sensor.
Biosensing Techniques , Nanostructures , Carbon/chemistry , Nanostructures/chemistry , Porosity , Silicon/chemistry
The biomolecular contents of extracellular vesicles, such as exosomes, have been shown to be crucial in intercellular communication and disease propagation. As a result, there has been a recent surge in the exploration of novel biosensing platforms that can sensitively and specifically detect exosomal content such as proteins and nucleic acids, with a view toward application in diagnostic assays. Here, we demonstrate dual-mode and label-free detection of plasma exosomes using an electrochemical quartz crystal microbalance with dissipation monitoring (EQCM-D). The platform adopts a direct immunosensing approach to effectively capture exosomes via their surface protein expression of CD63. By combining QCM-D with a tandem in situ electrochemical impedance spectroscopy measurement, we are able to demonstrate relationships between mass, viscoelasticity and impedance inducing properties of each functional layer and analyte. In addition to lowering the limit of detection (by a factor of 2-4) to 6.71 × 107 exosome-sized particles (ESP) per mL in 25% v/v serum, the synergy between dissipation and impedance response introduces improved sensing specificity by offering further distinction between soft and rigid analytes, thereby promoting EQCM-D as an important technique for exosome analysis.
Biosensing Techniques , Exosomes , Nucleic Acids , Biosensing Techniques/methods , Electric Impedance , Quartz , Quartz Crystal Microbalance Techniques/methods
Wearable technologies have great potential in health monitoring and disease diagnostics. As a consequence, interest in the study of wearable sensors has dramatically increased over recent years. Successful translation of this technology from research prototypes to commercial products requires addressing some of the major challenges faced by wearable sensors such as loss of, and damage in, the biological recognition layer of the skin-interfaced sensors. In this work, we propose a solution to this challenge by integrating micropillar array (MPA) surfaces as part of the sensing layer with the aim to protect and prevent the loss of the enzyme layer from mechanical stress while the sensor is worn. The proposed wearable sensing patch is composed of reference, counter, and working electrodes, all made of MPAs and is designed for measuring glucose in sweat. MPA sensing patch has a wide linear range of 50 µM to 1.4 mM, a sensitivity of 4.7 ± 0.8 µA mM-1, and a limit of detection of 26 ± 5 µM. The glucose sensing patch was tested using human sweat where glucose-level changes were successfully measured before and after meal consumption. The developed patch provides an alternative solution to the problem of the damage to the sensor microenvironment upon wear. But in addition, it also offers a user-friendly, cost-effective, and reliable sweat analysis platform with significant potential in health monitoring applications.
Biosensing Techniques , Glucose/analysis , Microfluidic Analytical Techniques , Silicon/chemistry , Wearable Electronic Devices , Humans , Surface Properties
Electrochemical devices for transdermal monitoring of key biomarkers are the potential next frontier of wearable technologies for point-of-care disease diagnosis, including Cancer in which Cancer is the leading cause of death worldwide with estimated 10 million deaths in 2018 according to the World Health Organization and breast cancer is one of the five most common causes of cancer death with over two million cases recorded in 2018. Early diagnosis and prognosis based on monitoring of breast cancer biomarkers is of high importance. In this work, high-density gold coated silicon microneedle arrays (Au-Si-MNA) were simultaneously used as biomarker extraction platform and electrochemical transducer, enabling the selective immunocapture of epidermal growth factor receptor 2 (ErbB2), a key breast cancer biomarker, and its subsequent quantification. The analytical performance of the device was tested in artificial interstitial fluid exhibiting a linear response over a wide concentration range from 10 to 250 ng/mL, with a detection limit of 4.8 ng/mL below the biomarker levels expected in breast cancer patients. As a proof of concept, the immunosensor demonstrated its ability to successfully extract ErbB2 from a phantom gel mimicking the epidermis and dermis layers, and subsequently quantify it showing a linear range from 50 to 250 ng/mL and a detection limit of 25 ng/mL. The uniqueness of this sensing platform combining direct transdermal biomarker extraction and quantification opens up new avenues towards the development of high performing wearable point-of-care devices.
Biosensing Techniques , Breast Neoplasms , Metal Nanoparticles , Biomarkers, Tumor , Breast Neoplasms/diagnosis , Electrochemical Techniques , Female , Gold , Humans , Immunoassay , Limit of Detection , Silicon
Porous polymeric membranes have shown great potential in biological and biomedical applications such as tissue engineering, bioseparation, and biosensing, due to their structural flexibility, versatile surface chemistry, and biocompatibility. This review outlines the advantages and limitations of the fabrication techniques commonly used to produce porous polymeric membranes, with especial focus on those featuring nano/submicron scale pores, which include track etching, nanoimprinting, block-copolymer self-assembly, and electrospinning. Recent advances in membrane technology have been key to facilitate precise control of pore size, shape, density and surface properties. The review provides a critical overview of the main biological and biomedical applications of these porous polymeric membranes, especially focusing on drug delivery, tissue engineering, biosensing, and bioseparation. The effect of the membrane material and pore morphology on the role of the membranes for each specific application as well as the specific fabrication challenges, and future prospects of these membranes are thoroughly discussed.
Biocompatible Materials/chemistry , Membranes, Artificial , Polymers/chemistry , Animals , Humans , Porosity
Nanotechnology's rapid development of nanostructured materials with disruptive material properties has inspired research for their use as electrocatalysts to potentially substitute enzymes. Herein, a novel electrocatalytic nanomaterial was constructed by growing gold nanograss (AuNG) on 2D nanoassemblies of gold nanocubes (AuNC). The resulting structure (NG@NC) was used for the detection of H2O2via its electrochemical reduction. The NG@NC electrode displayed a large active surface area, resulting in improved electron transfer efficiency. On the nanoscale, AuNG maintained its structure, providing high stability and reproducibility of the sensing platform. Our nanostructured electrode showed excellent catalytic activity towards H2O2 at an applied potential of -0.5 V vs Ag/AgCl. This facilitated H2O2 detection with excellent selectivity in an environment like human urine, and a linear response from 50 µM to 30 mM, with a sensitivity of 100.66 ± 4.0 µA mM-1 cm-2. The NG@NC-based sensor hence shows great potential in nonenzymatic electrochemical sensing.
A label-free electrochemical detection platform for the sensitive and rapid detection of Flightless I (Flii) protein, a biomarker of wound chronicity, has been developed using nanoporous anodic alumina (NAA) membranes modified with Flii antibody recognition sites. The electrochemical detection is based on the nanochannel blockage experienced upon Flii capture by immobilized antibodies within the nanochannels. This capture impedes the diffusion of redox species [[Fe(CN)6]4-/3-] toward a gold electrode attached at the backside of the modified NAA membrane. Partial blockage causes a decrease in the oxidation current of the redox species at the electrode surface which is used as an analytical signal by the reported biosensor. The resulting biosensing system allows detection of Flii at the levels found in wounds. Two types of assays were tested, sandwich and direct, showing <3 and 2 h analysis time, respectively, a significant reduction in time from the nearly 48 h required for the conventional Western blot assay. Slightly higher sensitivity values were observed for the sandwich-based strategy. With faster analysis, lack of matrix effects, robustness, ease of use and cost-effectiveness, the developed sensing platform has the potential to be translated into a point-of-care (POC) device for chronic wound management and as a simple alternative characterization tool in Flii research.
We report the first method to introduce differential functionalities in the interior pore walls and exterior surface of highly stable thermally hydrocarbonised porous silicon (THCpSi) films. The approach exploits the hydrophobicity of the hydrosilylated THCpSi to, first, selectively functionalise the external surface, and subsequently derivatise the hydrophobic internal pore walls.
A novel pore blockage-based electrochemical immunosensor based on the combination of 100 nm-magnetic nanoparticles (MNPs), as signal enhancers, and 200 nm-pore diameter nanoporous anodic alumina (NAA) membranes, as sensing platform, is reported. A peptide conjugate mimicking flightless I (Flii), a wound healing biomarker, was chosen as target analyte. The sensing platform consists of an anti-Flii antibody (Ab1)-modified NAA membrane attached onto a gold electrode. Anti-KLH antibody (Ab2)-modified MNPs (MNP-Ab2) were used to selectively capture the Flii peptide conjugate in solution. Sensing was based on pore blockage of the Ab1-modified NAA membrane caused upon specific binding of the MNP-Ab2-analyte complex. The degree of pore blockage, and thus the concentration of the Flii peptide conjugate in the sample, was measured as a reduction in the oxidation current of a redox species ([Fe(CN)6]4-) added in solution. We demonstrated that pore blockage is drastically enhanced by applying an external magnetic field at the membrane backside to facilitate access of the MNP-Ab2-analyte complex into the pores, and thus ensure its availability to bind to the Ab1-modified NAA membrane. Combining the pore blockage-based electrochemical magnetoimmunosensor with an externally applied magnetic field, a limit of detection (LOD) of 0.5 ng/ml of Flii peptide conjugate was achieved, while sensing in the absence of magnetic field could only attain a LOD of 1.2 µg/ml. The developed sensing strategy is envisaged as a powerful solution for the ultra-sensitive detection of an analyte of interest present in a complex matrix.
Lipopolysaccharides (LPS) are the major component of the outer membrane of all Gram-negative bacteria and some cyanobacteria and are released during growth and cell death. LPS pose a potential health risk in water, causing acute respiratory illnesses, inhalation fever, and gastrointestinal disorders. The need for rapid and accurate detection of LPS has become a major priority to facilitate more timely and efficacious intervention and, hence, avoid unsafe water distribution. In this context, a porous silicon membrane (pSiM)-based electrochemical biosensor was developed for direct and sensitive detection of LPS. pSiM, featuring arrays of nanochannels, was modified with polymyxin B (PmB), an antimicrobial peptide with strong affinity to LPS. Detection of LPS was based on measuring the changes in the diffusion through the nanochannels of an electroactive species added in solution, caused by the nanochannel blockage upon LPS binding to PmB. Results showed a limit of detection of 1.8 ng/mL, and a linear response up to 10,000 ng/mL spiked in buffer. Selectivity of the sensor toward potential interfering species in water supplies was also assessed. Sensor performance was then evaluated in water samples from a water treatment plant (WTP), and detection of LPS well below the levels encountered in episodes of water contamination and in humidifiers was demonstrated. The same platform was also tested for bacterial detection including Pseudomonas aeruginosa and Escherichia coli spiked in water samples from a WTP. Considering its performance characteristics, this platform represents a promising screening tool to identify the presence of LPS in water supplies and provide early warning of contamination events.
Biosensing Techniques/methods , Electrochemical Techniques/methods , Lipopolysaccharides/analysis , Water Microbiology , Dental Cements/chemistry , Electrochemical Techniques/instrumentation , Electrodes , Escherichia coli/chemistry , Lipopolysaccharides/metabolism , Membranes, Artificial , Polymyxin B/chemistry , Polymyxin B/metabolism , Pseudomonas aeruginosa/chemistry , Salmonella typhimurium/chemistry
