Lateral Flow Assay Biotesting by Utilizing Plasmonic Nanoparticles Made of Inexpensive Metals─Replacing Colloidal Gold.

Nanoparticles (NPs) can be conjugated with diverse biomolecules and employed in biosensing to detect target analytes in biological samples. This proven concept was primarily used during the COVID-19 pandemic with gold-NP-based lateral flow assays (LFAs). Considering the gold price and its worldwide depletion, here we show that novel plasmonic NPs based on inexpensive metals, titanium nitride (TiN) and copper covered with a gold shell (Cu@Au), perform comparable to or even better than gold nanoparticles. After conjugation, these novel nanoparticles provided high figures of merit for LFA testing, such as high signals and specificity and robust naked-eye signal recognition. Since the main cost of Au NPs in commercial testing kits is the colloidal synthesis, our development with the Cu@Au and the laser-ablation-fabricated TiN NPs is exciting, offering potentially inexpensive plasmonic nanomaterials for various bioapplications. Moreover, our machine learning study showed that biodetection with TiN is more accurate than that with Au.

Impedance-Based Detection of Bacteria.

Pathogenic bacteria have always posed one of the most serious threats to public health, and continue to be especially dangerous with the rise in antibiotic resistance. The prevalence of these infectious agents necessitates rapid, point-of-care sensors for their detection, identification, and monitoring. Electrochemical sensors are promising for the low-cost monitoring of bacterial growth and the detection of specific microbial species due to the consistency and ease-of-use of impedance measurements. Though the commercialization of these sensors is currently limited, they offer significant promise for detecting pathogens from real-world environments.

Electrochemically Controlled ATRP for Cleavage-Based Electrochemical Detection of the Prostate-Specific Antigen at Femtomolar Level Concentrations.

As a single-chain glycoprotein with endopeptidase activity, the prostate-specific antigen (PSA) is valuable as an informative serum marker in diagnosing, staging, and prognosis of prostate cancer. In this report, an electrochemical biosensor based on the target-induced cleavage of a specific peptide substrate (PSA peptide) is designed for the highly selective detection of PSA at the femtomolar level, using electrochemically controlled atom transfer radical polymerization (eATRP) as a method for signal amplification. The PSA peptides, without free carboxyl sites, are attached to the gold surface via the N-terminal cysteine residue. The target-induced cleavage of PSA peptides results in the generation of carboxyl sites, to which the alkyl halide initiator α-bromophenylacetic acid (BPAA) is linked via the Zr(IV) linkers. Subsequently, the potentiostatic eATRP of ferrocenylmethyl methacrylate (FcMMA, as the monomer) leads to the surface-initiated grafting of high-density ferrocenyl polymers. As a result, a large amount of Fc redox tags can be recruited for signal amplification, through which the limit of detection (LOD) for PSA can be down to 3.2 fM. As the recognition element, the PSA peptide is easy to synthesize, chemically and thermally stable, and low-cost. Without the necessity of enzyme or nanoparticle labels, the eATRP-based amplification method is easy to operate and low-cost. Results also show that the cleavage-based electrochemical PSA biosensor is highly selective and applicable to PSA detection in complex biological samples. In view of these merits, the integration of the eATRP-based amplification method into cleavage-based recognition is believed to hold great promise for the electrochemical detection of PSA in clinical applications.

Performance limitations of resonant refractive index sensors with low-cost components.

Resonant biosensors are attractive for diagnostics because they can detect clinically relevant biomarkers with high sensitivity and in a label-free fashion. Most of the current solutions determine their detection limits in a highly stabilised laboratory environment, which does, however, not apply to real point-of-care applications. Here, we consider the more realistic scenario of low-cost components and an unstabilised environment and consider the related design implications. We find that sensors with lower quality-factor resonances are more fault tolerant, that a filtered LED lightsource is advantageous compared to a diode laser, and that a CMOS camera is preferable to a CCD camera for detection. We exemplify these findings with a guided mode resonance sensor and experimentally determine a limit of detection of 5.8 ± 1.7×10-5 refractive index units (RIU), which is backed up by a model identifying the various noise sources. Our findings will inform the design of high performance, low cost biosensors capable of operating in a real-world environment.

A low-cost paper-based aptasensor for simultaneous trace-level monitoring of mercury (II) and silver (I) ions.

Mercury (Hg2+) and silver (Ag+) ions possess the harmful effects on public health and environment that makes it essential to develop the sensing techniques with great sensitivity for the ions. Metal ions commonly coexist in the different biological and environmental systems. Hence, it is an urgent demand to design a simple method for the simultaneous detection of metal ions, peculiarly in the case of coexisting Hg2+ and Ag+. This study introduces a low-cost paper-based aptasensor to monitor Hg2+ and Ag+, simultaneously. The strategy of the sensing array is according to the conformational changes of Hg2+- and Ag+-specific aptamers and their release from the GO surface after the injection of the target sample on the sensing platform. Through monitoring the fluorescence recovery changes against the concentrations of the ions, Hg2+ and Ag+ can be determined as low as 1.33 and 1.01 pM. The paper-based aptasensor can simultaneously detect the ions within about 10 min. The aptasensor is applied prosperously to monitor Hg2+ and Ag+ in human serum, water, and milk. The designed aptasensor with the main advantages of simplicity and feasibility holds the supreme potential to develop a cost-effective sensing method for environmental monitoring, food control, and human diagnostics.

A Cost-Effective and Portable Optical Sensor System to Estimate Leaf Nitrogen and Water Contents in Crops.

Non-invasive determination of leaf nitrogen (N) and water contents is essential for ensuring the healthy growth of the plants. However, most of the existing methods to measure them are expensive. In this paper, a low-cost, portable multispectral sensor system is proposed to determine N and water contents in the leaves, non-invasively. Four different species of plants-canola, corn, soybean, and wheat-are used as test plants to investigate the utility of the proposed device. The sensor system comprises two multispectral sensors, visible (VIS) and near-infrared (NIR), detecting reflectance at 12 wavelengths (six from each sensor). Two separate experiments were performed in a controlled greenhouse environment, including N and water experiments. Spectral data were collected from 307 leaves (121 for N and 186 for water experiment), and the rational quadratic Gaussian process regression (GPR) algorithm was applied to correlate the reflectance data with actual N and water content. By performing five-fold cross-validation, the N estimation showed a coefficient of determination () of 63.91% for canola, 80.05% for corn, 82.29% for soybean, and 63.21% for wheat. For water content estimation, canola showed an of 18.02%, corn showed an of 68.41%, soybean showed an of 46.38%, and wheat showed an of 64.58%. The result reveals that the proposed low-cost sensor with an appropriate regression model can be used to determine N content. However, further investigation is needed to improve the water estimation results using the proposed device.

A Novel Method to Estimate the Full Knee Joint Kinematics Using Low Cost IMU Sensors for Easy to Implement Low Cost Diagnostics.

Traditional motion capture systems are the current standard in the assessment of knee joint kinematics. These systems are, however, very costly, complex to handle, and, in some conditions, fail to estimate the varus/valgus and internal/external rotation accurately due to the camera setup. This paper presents a novel and comprehensive method to infer the full relative motion of the knee joint, including the flexion/extension, varus/valgus, and internal/external rotation, using only low cost inertial measurement units (IMU) connected to the upper and lower leg. Furthermore, sensors can be placed arbitrarily and only require a short calibration, making it an easy-to-use and portable clinical analysis tool. The presented method yields both adequate results and displays the uncertainty band on those results to the user. The proposed method is based on an fixed interval smoother relying on a simple dynamic model of the legs and judicially chosen constraints to estimate the rigid body motion of the leg segments in a world reference frame. In this pilot study, benchmarking of the method on a calibrated robotic manipulator, serving as leg analogue, and comparison with camera-based techniques confirm the method's accurateness as an easy-to-implement, low-cost clinical tool.

An Intact Cell Bioluminescence-Based Assay for the Simple and Rapid Diagnosis of Urinary Tract Infection.

Urinary tract infection (UTI) is one of the most common infections, accounting for a substantial portion of outpatient hospital and clinic visits. Standard diagnosis of UTI by culture and sensitivity can take at least 48 h, and improper diagnosis can lead to an increase in antibiotic resistance following therapy. To address these shortcomings, rapid bioluminescence assays were developed and evaluated for the detection of UTI using intact, viable cells of Photobacterium mandapamensis USTCMS 1132 or previously lyophilized cells of Photobacterium leiognathi ATCC 33981™. Two platform technologies-tube bioluminescence extinction technology urine (TuBETUr) and cellphone-based UTI bioluminescence extinction technology (CUBET)-were developed and standardized using artificial urine to detect four commonly isolated UTI pathogens-namely, Escherichia coli, Proteus mirabilis, Staphylococcus aureus, and Candida albicans. Besides detection, these assays could also provide information regarding pathogen concentration/level, helping guide treatment decisions. These technologies were able to detect microbes associated with UTI at less than 105 CFU/mL, which is usually the lower cut-off limit for a positive UTI diagnosis. Among the 29 positive UTI samples yielding 105-106 CFU/mL pathogen concentrations, a total of 29 urine specimens were correctly detected by TuBETUr as UTI-positive based on an 1119 s detection window. Similarly, the rapid CUBET method was able to discriminate UTIs from normal samples with high confidence (p ≤ 0.0001), using single-pot conditions and cell phone-based monitoring. These technologies could potentially address the need for point-of-care UTI detection while reducing the possibility of antibiotic resistance associated with misdiagnosed cases of urinary tract infections, especially in low-resource environments.

Facile and Low-Cost SPE Modification Towards Ultra-Sensitive Organophosphorus and Carbamate Pesticide Detection in Olive Oil.

Despite the fact that a considerable amount of effort has been invested in the development of biosensors for the detection of pesticides, there is still a lack of a simple and low-cost platform that can reliably and sensitively detect their presence in real samples. Herein, an enzyme-based biosensor for the determination of both carbamate and organophosphorus pesticides is presented that is based on acetylcholinesterase (AChE) immobilized on commercially available screen-printed carbon electrodes (SPEs) modified with carbon black (CB), as a means to enhance their conductivity. Most interestingly, two different methodologies to deposit the enzyme onto the sensor surfaces were followed; strikingly different results were obtained depending on the family of pesticides under investigation. Furthermore, and towards the uniform application of the functionalization layer onto the SPEs' surfaces, the laser induced forward transfer (LIFT) technique was employed in conjunction with CB functionalization, which allowed a considerable improvement of the sensor's performance. Under the optimized conditions, the fabricated sensors can effectively detect carbofuran in a linear range from 1.1 × 10-9 to 2.3 × 10-8 mol/L, with a limit of detection equal to 0.6 × 10-9 mol/L and chlorpyrifos in a linear range from 0.7 × 10-9 up to 1.4 × 10-8 mol/L and a limit of detection 0.4 × 10-9 mol/L in buffer. The developed biosensor was also interrogated with olive oil samples, and was able to detect both pesticides at concentrations below 10 ppb, which is the maximum residue limit permitted by the European Food Safety Authority.

Project honeybee: Clinical applications for wearable biosensors.

Please provide an abstract of 150 to 250 words. The abstract should not contain any undefined abbreviations or unspecified references. The Project Honeybee Observational Clinical Trials were 12-month studies designed to validate the use of commercially available ambulatory medical devices costing $50-$300 for clinical applications. Each trial had a patient population of about 15-30 subjects with a broad range of disease types including heart failure, diabetes, sepsis, and Parkinson's disease. Over 30 supported proposals were funded in the 4-year period, as well as the creation of a database of all commercially available devices. Each year a call for proposals was published within ASU and Mayo Clinic Arizona. Proposals were selected for funding by a committee of ASU faculty from engineering, nursing, and exercise physiology departments. The progress of each research trial was monitored through monthly colloquia with the nursing, biomedical engineering, computer science, and nutrition graduate research assistants, to discuss the challenges and opportunities arising with each research trial. PIs were required to report on study progress 6 months into the trial period and 3 months following the conclusion of the 12-month project. The project was very successful in meeting our goals of testing consumer wearable devices on patients for a variety of conditions across a variety of clinical settings in the greater Phoenix community. The following clinical sites participated in one or more of these clinical trials: Adelante Healthcare, Arizona Arrhythmia Consultants, Arizona Cardiology Group, Banner University Medical Center, Barrow Neurological Institute, Honor Health, Mayo Clinic, and St Joseph's Hospital. A total of 12 ASU faculty and 39 clinicians participated.

Advances in immunosensors for clinical applications.

Immunoassay technique performs a fast, simple, reliable, and sensitive analysis of different compounds, being applied in several areas of interest such as clinical analysis for medical diagnosis, as well as in environmental analysis, and food quality control. The latest research activities in this field are represented by the attempts to achieve a low limit of detection by developing of new signal amplification strategies, eliminate the interferences, and decrease the cost of analysis.

Demonstration of a Low-Cost and Portable Optical Cavity-Based Sensor through Refractive Index Measurements.

An optical cavity-based sensor using a differential detection method has been proposed for point-of-care diagnostics. We developed a low-cost and portable optical cavity-based sensor system using a 3D printer and off-the-shelf optical components. In this paper, we demonstrate the sensing capability of the portable system through refractive index measurements. Fabricated optical cavity samples were tested using the portable system and compared to simulation results. A referencing technique and digital low pass filtering were applied to reduce the noise of the portable system. The measurement results match the simulation results well and show the improved linearity and sensitivity by employing the differential detection method. The limit of detection achieved was 1.73 × 10-5 Refractive Index Unit (RIU), which is comparable to other methods for refractive index sensing.

What are the Main Sensor Methods for Quantifying Pesticides in Agricultural Activities? A Review.

In recent years, there has been an increase in pesticide use to improve crop production due to the growth of agricultural activities. Consequently, various pesticides have been present in the environment for an extended period of time. This review presents a general description of recent advances in the development of methods for the quantification of pesticides used in agricultural activities. Current advances focus on improving sensitivity and selectivity through the use of nanomaterials in both sensor assemblies and new biosensors. In this study, we summarize the electrochemical, optical, nano-colorimetric, piezoelectric, chemo-luminescent and fluorescent techniques related to the determination of agricultural pesticides. A brief description of each method and its applications, detection limit, purpose-which is to efficiently determine pesticides-cost and precision are considered. The main crops that are assessed in this study are bananas, although other fruits and vegetables contaminated with pesticides are also mentioned. While many studies have assessed biosensors for the determination of pesticides, the research in this area needs to be expanded to allow for a balance between agricultural activities and environmental protection.

Programmable Nucleic Acid Nanoswitches for the Rapid, Single-Step Detection of Antibodies in Bodily Fluids.

Antibody detection plays a pivotal role in the diagnosis of pathogens and monitoring the success of vaccine immunization. However, current serology techniques require multiple, time-consuming washing and incubation steps, which limit their applicability in point-of-care (POC) diagnostics and high-throughput assays. We developed here a nucleic acid nanoswitch platform able to instantaneously measure immunoglobulins of type G and E (IgG and IgE) levels directly in blood serum and other bodily fluids. The system couples the advantages of target-binding induced colocalization and nucleic acid conformational-change nanoswitches. Due to the modular nature of the recognition platform, the method can potentially be applied to the detection of any antibody for which an antigen can be conjugated to a nucleic acid strand. In this work we show the sensitive, fast and cost-effective detection of four different antibodies and demonstrate the possible use of this approach for the monitoring of antibody levels in HIV+ patients immunized with AT20 therapeutic vaccine.

Antibody-Mediated Small Molecule Detection Using Programmable DNA-Switches.

The development of rapid, cost-effective, and single-step methods for the detection of small molecules is crucial for improving the quality and efficiency of many applications ranging from life science to environmental analysis. Unfortunately, current methodologies still require multiple complex, time-consuming washing and incubation steps, which limit their applicability. In this work we present a competitive DNA-based platform that makes use of both programmable DNA-switches and antibodies to detect small target molecules. The strategy exploits both the advantages of proximity-based methods and structure-switching DNA-probes. The platform is modular and versatile and it can potentially be applied for the detection of any small target molecule that can be conjugated to a nucleic acid sequence. Here the rational design of programmable DNA-switches is discussed, and the sensitive, rapid, and single-step detection of different environmentally relevant small target molecules is demonstrated.

Low-Cost and Rapid-Production Microfluidic Electrochemical Double-Layer Capacitors for Fast and Sensitive Breast Cancer Diagnosis.

This technical note describes a new microfluidic sensor that combines low-cost (USD $0.97) with rapid fabrication and user-friendly, fast, sensitive, and accurate quantification of a breast cancer biomarker. The electrodes consisted of cost-effective bare stainless-steel capillaries, whose mass production is already well-established. These capillaries were used as received, without any surface modification. Microfluidic chips containing electrical double-layer capillary capacitors (µEDLC) were obtained by a cleanroom-free prototyping that allows the fabrication of dozens to hundreds of chips in 1 h. This sensor provided the successful quantification of CA 15-3, a biomarker protein for breast cancer, in serum samples from cancer patients. Antibody-anchored magnetic beads were utilized for immunocapture of the marker, and then, water was added to dilute the protein. Next, the CA 15-3 detection (<2 min) was made without using redox probes, antibody on electrode (sandwich immunoassay), or signal amplification strategies. In addition, the capacitance tests eliminated external pumping systems and precise volumetric sampling steps, as well as presented low sample volume (5 µL) and high sensitivity using bare capillaries in a new design for double-layer capacitors. The achieved limit-of-detection (92.0 µU mL-1) is lower than that of most methods reported in the literature for CA 15-3, which are based on nanostructured electrodes. The data shown in this technical note support the potential of the µEDLC toward breast cancer diagnosis even at early stages. We believe that accurate analyses using a simple sample pretreatment such as magnetic field-assisted immunocapture and cost-effective bare electrodes can be extended to quantify other cancer biomarkers and even biomolecules by changing the biorecognition element.

Ultrafast Multiplexed-Allergen Detection through Advanced Fluidic Design and Monolithic Interferometric Silicon Chips.

A silicon-based miniaturized sensor chip combined with an advanced microfluidic module for the simultaneous, label-free immunochemical determination of four allergens, bovine milk protein, peanut protein, soy protein, and gliadin, is presented. The sensor chip consists of an array of 10 broad-band Mach-Zehnder interferometers (BB-MZIs) monolithically integrated on silicon, along with their respective broad-band light sources. The BB-MZIs were biofunctionalized with the targeted allergens and their responses during immunoreaction were monitored by multiplexing their transmission spectra through an external miniaturized spectrometer. The assay is performed by running mixtures of calibrators or samples with the antibodies against the four allergens followed by an antispecies specific antibodies solution. Employing a fluidic module of nearly one-dimensional geometry, that provided for uniform delivery of the reagents, CV values <6% were achieved for the responses of the 10 BB-MZIs, allowing for reliable multianalyte determinations. The analysis is completed in 6.5 min, and the detection limits were 0.04 µg/mL for bovine k-casein, 1.0 µg/mL for peanut protein, 0.80 µg/mL for soy protein, and 0.10 µg/mL for gliadin. The assays were accurate (recoveries 88-118%) and repeatable (intra- and interassay CVs <7% for all four allergens). Finally, the sensor was evaluated by analyzing samples from a cleaning in place system (CIP) of a dairy industry and the results obtained were in good agreement with those received by the respective ELISAs. The analytical characteristics of the sensor combined with the short analysis time and the small chip size make the proposed system an ideal tool for on-site multianalyte determinations.

Label-free real-time detection of biotinylated bovine serum albumin using a low-cost optical cavity-based biosensor.

We have developed a low-cost optical cavity-based biosensor with a differential detection method for point-of-care medical diagnostics. To experimentally demonstrate its label-free real-time biosensing capability, we performed the detection of biotinylated bovine serum albumin (BSA). Streptavidin is introduced into the optical cavity structure and immobilized on 3-aminopropyltriethoxysilane (APTES) coated surface. After rinsing out unbound streptavidin with DI water, biotinylated BSA without any labeling is introduced. A CMOS camera captures the transmitted light of two different wavelengths passing through the optical cavity sensing area in real-time. Then, the differential values are calculated to enhance the responsivity. We successfully demonstrated the label-free real-time detection of biotinylated BSA, and the measurement results matched well with the simulation results. The limit of detection of the optical cavity-based biosensor for the biotinylated BSA detection with the sensing area of 180 µm × 180 µm is estimated to be 2.82 pM, which could be reduced further for a smaller sensing area with the tradeoff of a longer sensing time.

Preparation of gold nanoparticles supported on graphene oxide with flagella as the template for nonenzymatic hydrogen peroxide sensing.

Gold nanoparticles supported on graphene oxide with flagella as the template were developed as an electrochemical sensor for the detection of hydrogen peroxide (H2O2) in serum. The flagella-Au nanoparticles composite and graphene oxide were dropped onto a glassy carbon electrode (GCE) to form a new H2O2 electrochemical sensor. The structure morphology of the prepared sensor was characterized by transmission electron microscopy (TEM), and the electrocatalytic performance towards H2O2 reduction was evaluated by cyclic voltammetry (CV) and amperometric methods. The response current of the sensor showed a good linear relationship with the concentration of H2O2 in the range of 10-1000 µM (R2 = 0.9916). The minimum detection limit of 1 µM was obtained (S/N = 3). Finally, the sensor was applied to the detection of H2O2 in serum, and the recoveries were satisfactory. As the sensor is sensitive, fast, and easy to make, it is expected to be used for rapid detection of H2O2. Graphical abstract ᅟ.

Rapid recognition of volatile organic compounds with colorimetric sensor arrays for lung cancer screening.

Volatile organic compounds (VOCs) in breath can be used as biomarkers to identify early stages of lung cancer. Herein, we report a disposable colorimetric array that has been constructed from diverse chemo-responsive colorants. Distinguishable difference maps were plotted within 4 min for specifically targeted VOCs. Through the consideration of various chemical interactions with VOCs, the arrays successfully discriminate between 20 different volatile organic compounds in breath that are related to lung cancer. VOCs were identified either with the visualized difference maps or through pattern recognition with an accuracy of at least 90%. No uncertainties or errors were observed in the hierarchical cluster analysis (HCA). Finally, good reproducibility and stability of the array was achieved against changes in humidity. Generally, this work provides fundamental support for construction of simple and rapid VOC sensors. More importantly, this approach provides a hypothesis-free array method for breath testing via VOC profiling. Therefore, this small, rapid, non-invasive, inexpensive, and visualized sensor array is a powerful and promising tool for early screening of lung cancer. Graphical abstract A disposable colorimetric array has been developed with broadly chemo-responsive dyes to incorporate various chemical interactions, through which the arrays successfully discriminate 20 VOCs that are related to lung cancer via difference maps alone or chemometrics within 4 min. The hydrophobic porous matrix provides good stability against changes in humidity.