Ara H1 peanut allergen detection using a labelled electrochemical aptasensor based on GO-COOH@bimetallic composite platform.

Allergies are defined as a hypersensitivity reaction, immunologically mediated, as a result to an external stimulus. Peanuts induced allergies are considered one of the most severe, life-threatening food sensitivities since they trigger the highest frequency of severe and fatal reactions, even in trace amounts. Therefore, it is imperative to develop fast, accurate and easy-to-use analytical methods to determine Ara h1, is a seed storage protein from Arachis hypogea and the main peanut derived allergen. In this work, two strategies were applied to develop an electrochemical aptasensor based on GO-COOH and metallic nanoparticles immobilised on screen-printed carbon electrodes (SPCEs). The analytical performances of the aptasensor showed a linear range of 5-150 nM, and a limit of detection of 1.66 nM. The method was applied in peanut-free food samples with very good recoveries proving to be a promising tool for peanut allergy prevention.

Point-of-care electrochemical testing of biomarkers involved in inflammatory and inflammatory-associated medical conditions.

Recent years have shown that the diagnosis and monitoring of biomarkers involved in inflammatory-associated medical conditions such as cancer, neurological disorders, viral infections, or daily physical activities offer real benefits in increasing the quality of medical care and patient life quality. In this context, the use of integrated and portable platforms as point-of-care testing devices for biomedical analysis to enable early disease diagnosis and monitoring, which can be successfully used even at the patient's bed, is an emergency nowadays. The development of low-cost, miniaturized, and portable, user-friendly devices that provide an answer in a timely manner, such as electrochemical sensors, is relevant for the elaboration of point-of-care testing devices. This review focuses on the recent progress in bioanalysis of both specific biomarkers and inflammatory-associated biomarkers present in several diseases like neoplasia, severe neurological disorders, viral infections, and usual physical activity and provides an overview of the state of the art over the most recent electrochemical (bio)sensors for the detection of inflammation-related biomarkers. Future perspectives of point-of-care testing to improve healthcare management are also discussed.

Label-Free Electrochemical Aptasensor for the Detection of the 3-O-C12-HSL Quorum-Sensing Molecule in Pseudomonas aeruginosa.

Pseudomonas aeruginosa, an opportunistic Gram-negative bacterium, is one of the main sources of infections in healthcare environments, making its detection very important. N-3-oxo-dodecanoyl L-homoserine lactone (3-O-C12-HSL) is a characteristic molecule of quorum sensing-a form of cell-to-cell communication between bacteria-in P. aeruginosa. Its detection can allow the determination of the bacterial population. In this study, the development of the first electrochemical aptasensor for the detection of 3-O-C12-HSL is reported. A carbon-based screen-printed electrode modified with gold nanoparticles proved to be the best platform for the aptasensor. Each step in the fabrication of the aptasensor (i.e., gold nanoparticles' deposition, aptamer immobilization, incubation with the analyte) was optimized and characterized using cyclic voltammetry, differential pulse voltammetry, and electrochemical impedance spectroscopy. Different redox probes in solution were evaluated, the best results being obtained in the presence of [Fe(CN)6]4-/[Fe(CN)6]3-. The binding affinity of 106.7 nM for the immobilized thiol-terminated aptamer was determined using surface plasmon resonance. The quantification of 3-O-C12-HSL was performed by using the electrochemical signal of the redox probe before and after incubation with the analyte. The aptasensor exhibited a logarithmic range from 0.5 to 30 µM, with a limit of detection of 145 ng mL-1 (0.5 µM). The aptasensor was successfully applied for the analysis of real samples (e.g., spiked urine samples, spiked microbiological growth media, and microbiological cultures).

Analytical methods for the characterization and diagnosis of infection with Pseudomonas aeruginosa: A critical review.

The recent increase in outbreaks of pathogenic bacteria and antimicrobial resistance represent major public health problems. Being the leading cause of death in humans, the bacterial infections need to be accurately and quickly diagnosed. Hence, the development of fast, cost-effective, sensitive and specific strategies for the detection of the targeted bacterium is of utmost importance. This review presents a systematic, critical evaluation of the recent analytical methods for the characterization and diagnosis of infections caused by Pseudomonas aeruginosa. The clinical manifestations, incidence and treatment of the P. aeruginosa infection and the associated quorum sensing, biofilm formation and virulence factors are also discussed. An overview of a variety of analytical methods for the detection of P. aeruginosa is provided, including whole-cell detection (microbiological methods, biosensors), antigens, DNA, and relevant markers (quorum sensing molecules, virulence factors) detection. The latest trends in analytical methods, especially sensors, are the orientation towards portability and on-site detection. The efforts made so far to achieve these goals in the detection of P. aeruginosa and its markers are also presented and discussed in this review. The strengths and weaknesses of the current detection methods are evaluated, while exploring potential routes for further development.

Gold-based nanostructured platforms for oxytetracycline detection from milk by a "signal-on" aptasensing approach.

Several gold platforms of different morphologies were investigated in the elaboration of a new aptasensor for oxytetracycline. Au-nanostructures were electrochemically synthesized from solutions of different concentrations of HAuCl4 in different media by chronoamperometry, multipulse amperometry, and chronopotentiometry, respectively at carbon-based screen-printed electrodes (C-SPE). The nano-/micro-scale morphologies of the patterned surfaces and elemental composition were examined by scanning electron microscopy, atomic force microscopy, and X-ray photoelectron spectroscopy, respectively. The electrochemical properties of the obtained gold nanostructured platforms (AuNSs|C-SPE) were investigated to achieve optimal aptamer coverage. The results showed that the aptasensor developed using the platform with thistle-like AuNSs exhibited the highest conductivity in terms of ferrocene signal and the largest effective area. Under optimal conditions, a linear range from 5.0 × 10-8 M to 1.2 × 10-6 M, with a limit of detection (LOD) of 8.7 × 10-9 M OXT were obtained, which is about 20 times lower than the EU regulations for OXT residues in milk. The electrochemical aptasensor was able to discriminate other antibacterial agents, such as amoxicillin, ampicillin, gentamicin, tetracycline, and vancomycin and was successfully applied in milk samples. This "signal-on" aptasensing approach provides a simple and cost-effective disposable sensor that could be easily applied for the on-site determination of antibiotics.

Electrochemical Peptide-Based Sensors for Foodborne Pathogens Detection.

Food safety and quality control pose serious issues to food industry and public health domains, in general, with direct effects on consumers. Any physical, chemical, or biological unexpected or unidentified food constituent may exhibit harmful effects on people and animals from mild to severe reactions. According to the World Health Organization (WHO), unsafe foodstuffs are especially dangerous for infants, young children, elderly, and chronic patients. It is imperative to continuously develop new technologies to detect foodborne pathogens and contaminants in order to aid the strengthening of healthcare and economic systems. In recent years, peptide-based sensors gained much attention in the field of food research as an alternative to immuno-, apta-, or DNA-based sensors. This review presents an overview of the electrochemical biosensors using peptides as molecular bio-recognition elements published mainly in the last decade, highlighting their possible application for rapid, non-destructive, and in situ analysis of food samples. Comparison with peptide-based optical and piezoelectrical sensors in terms of analytical performance is presented. Methods of foodstuffs pretreatment are also discussed.

Hybrid Nanocomposite Platform, Based on Carbon Nanotubes and Poly(Methylene Blue) Redox Polymer Synthesized in Ethaline Deep Eutectic Solvent for Electrochemical Determination of 5-Aminosalicylic Acid.

A novel hybrid composite of conductive poly(methylene blue) (PMB) and carbon nanotubes (CNT) was prepared for the detection of 5-aminosalicylic acid (5-ASA). Electrosynthesis of PMB with glassy carbon electrode (GCE) or with carbon nanotube modified GCE was done in ethaline deep eutectic solvent of choline chloride mixed with ethylene glycol and a 10% v/v aqueous solution. Different sensor architectures were evaluated in a broad range of pH values in a Britton-Robinson (BR) buffer using electrochemical techniques, chronoamperometry (CA), and differential pulse voltammetry (DPV), to determine the optimum sensor configuration for 5-ASA sensing. Under optimal conditions, the best analytical performance was obtained with CNT/PMBDES/GCE in 0.04 M BR buffer pH 7.0 in the range 5-100 µM 5-ASA using the DPV method, with an excellent sensitivity of 9.84 µA cm-2 µM-1 (4.9 % RSD, n = 5) and a detection limit (LOD) (3σ/slope) of 7.7 nM, outclassing most similar sensors found in the literature. The sensitivity of the same sensor obtained in CA (1.33 µA cm-2 µM-1) under optimal conditions (pH 7.0, Eapp = +0.40 V) was lower than that obtained by DPV. Simultaneous detection of 5-ASA and its analogue, acetaminophen (APAP), was successfully realized, showing a catalytic effect towards the electro-oxidation of both analytes, lowering their oxidation overpotential, and enhancing the oxidation peak currents and peak-to-peak separation as compared with the unmodified electrode. The proposed method is simple, sensitive, easy to apply, and economical for routine analysis.

Tackling the Problem of Sensing Commonly Abused Drugs Through Nanomaterials and (Bio)Recognition Approaches.

We summarize herein the literature in the last decade, involving the use of nanomaterials and various (bio)recognition elements, such as antibodies, aptamers and molecularly imprinted polymers, for the development of sensitive and selective (bio)sensors for illicit drugs with a focus on electrochemical transduction systems. The use and abuse of illicit drugs remains an increasing challenge for worldwide authorities and, therefore, it is important to have accurate methods to detect them in seized samples, biological fluids and wastewaters. They are recently classified as the latest group of "emerging pollutants," as their consumption has increased tremendously in recent years. Nanomaterials, antibodies, aptamers and molecularly imprinted polymers have gained much attention over the last decade in the development of (bio)sensors for a myriad of applications. The applicability of these (nano)materials, functionalized or not, has significantly increased, and are therefore highly suitable for use in the detection of drugs. Lately, such functionalized nanoscale materials have assisted in the detection of illicit drugs fingerprints, providing large surface area, functional groups and unique properties that facilitate sensitive and selective sensing. The review discusses the types of commonly abused drugs and their toxicological implications, classification of functionalized nanomaterials (graphene, carbon nanotubes), their fabrication, and their application on real samples in different fields of forensic science. Biosensors for drugs of abuse from the last decade's literature are then exemplified. It also offers insights into the prospects and challenges of bringing the functionalized nanobased technology to the end user in the laboratories or in-field.

Colorimetric multienzymatic smart sensors for hydrogen peroxide, glucose and catechol screening analysis.

In this work, we present a smartphone-based multiplexed enzymatic biosensor utilizing the unique colorimetric properties of the poly(aniline-co-anthranilic acid) (ANI-co-AA) composite film coupled with horseradish peroxidase (HRP), glucose oxidase (GOx), horseradish peroxidase-glucose oxidase (GOx-HRP) and tyrosinase (Tyr) enzymes. The enzymes are immobilized on the composite polymer film by adsorption and they catalyze a reversible redox color change of the host polymer from green to blue in the presence of their substrate. A smartphone was applied as color detector, for image acquisition and data handling. A ColorLab® android application, free of charge software application, was used to enable easy and clear display of the sensors' response indicating remarkable changes in the optical features. The results were confirmed by the spectrophotometric measurements. The developed colorimetric enzymatic biosensors were studied and optimized in relation to different experimental parameters. Moreover, the colorimetric enzymatic biosensors were applied to food and pharmaceutical analysis. It has been shown by these studies that the colorimetric biosensors are promising as quick and simple tests for handheld analysis in various fields.

Latest Trends in Electrochemical Sensors for Neurotransmitters: A Review.

Neurotransmitters are endogenous chemical messengers which play an important role in many of the brain functions, abnormal levels being correlated with physical, psychotic and neurodegenerative diseases such as Alzheimer's, Parkinson's, and Huntington's disease. Therefore, their sensitive and robust detection is of great clinical significance. Electrochemical methods have been intensively used in the last decades for neurotransmitter detection, outclassing more complicated analytical techniques such as conventional spectrophotometry, chromatography, fluorescence, flow injection, and capillary electrophoresis. In this manuscript, the most successful and promising electrochemical enzyme-free and enzymatic sensors for neurotransmitter detection are reviewed. Focusing on the activity of worldwide researchers mainly during the last ten years (2010-2019), without pretending to be exhaustive, we present an overview of the progress made in sensing strategies during this time. Particular emphasis is placed on nanostructured-based sensors, which show a substantial improvement of the analytical performances. This review also examines the progress made in biosensors for neurotransmitter measurements in vitro, in vivo and ex vivo.

Electrochemical Fingerprint of Arsenic (III) by Using Hybrid Nanocomposite-Based Platforms.

Arsenic, one of the most abundant mineral and also one to the most toxic compounds. Due to its high toxicity sensitive analytical methods are highly important, taking into account that the admitted level is in the range of µg L-1. A novel and easy to use platform for As(III) detection from water samples is proposed, based on gold and platinum bi metallic nanoparticles and a conductive polymer (polyaniline). The electrochemical detection was achieved after optimization of cathodic pre-concentration and stripping parameters by square wave anodic stripping voltammetry at modified screen-printed carbon-based electrochemical cells, proving its applicability for disposable and cost-effective in situ analysis of arsenic.

DNA-Based Sensor for the Detection of an Organophosphorus Pesticide: Profenofos.

In this work, we propose an electrochemical DNA aptasensor for the detection of profenofos, an organophosphorus pesticide, based on a competitive format and disposable graphite screen-printed electrodes (GSPEs). A thiol-tethered DNA capture probe, which results to be complementary to the chosen aptamer sequence, was immobilised on gold nanoparticles/polyaniline composite film-modified electrodes (AuNPs/PANI/GSPE). Different profenofos solutions containing a fixed amount of the biotinylated DNA aptamer were dropped onto the realized aptasensors. The hybridisation reaction was measured using a streptavidin-alkaline phosphatase enzyme conjugate, which catalyses the hydrolysis of 1-naphthyl -phosphate. The 1-naphtol enzymatic product was detected by means of differential pulse voltammetry (DPV). The aptasensor showed itself to work as a signal off sensor, according to the competitive format used. A dose response curve was obtained between 0.10 μM and 10 μM with a detection limit of 0.27 μM.

Electrochemical Immunosensors for Disease Detection and Diagnosis.

BACKGROUND: The detection of biological molecules referred as biomarkers in biological fluids is fundamental in clinical analysis because it permits to discriminate between healthy and ill individuals and to evaluate the progress of a disease. The development of immunosensors for the detection and monitoring of biomarkers is currently a major area of research and, as more markers are discovered and their role in disease becomes better understood, this will continue to grow. METHODS: We report the research progresses of electrochemical immunosensor applied in clinical analysis that have been published in the last three years. RESULTS: The emphasis of this review is on the advances of the electrochemical immunosensors for detection and monitoring of cancerous, cardiovascular and neurological diseases. An immunosensor overview was presented as well as the biomarkers and biosensing systems currently used to detect the onset and monitor the progression of the mentioned diseases. CONCLUSIONS: Electrochemical biosensors focusing on a vast repertoire of analytes are now becoming one of the most widely explored scientific fields. This is due to their enormous potential in clinical diagnosis and biological process monitoring. In the near future, with the development of transducer technology, nano-sized material technology, and biomolecules engineering technology, biosensors should be powerful tools in several analytical areas.

Smartphone-based immunosensor for CA125 detection.

In this work, we report the design, the development and the characterization of the analytical performances of a colorimetric smartphone-based immunosensor for the detection of cancer antigen 125 (CA125). The immunosensor was based on a sandwich strategy in which the primary antibody was immobilized by spotting onto the 3D nitrocellulose membrane. The immunospots were subsequently incubated with CA125 solutions, followed by the affinity reaction with a secondary antibody labeled with gold nanoparticles (AuNPs). The antibody-AuNPs captured onto immunospots induced the silver deposition from a silver enhancer solution leading to the formation of gold-silver nanoparticles of different grey color spots depending on CA125 concentration. The 8 megapixels smartphone camera was integrated in a home-made dark box and used as transducer of color image acquisition and data handling. The pixel intensity of the captured images was determined by an image processing algorithm. The experimental parameters involved in each step of the immunosensor design were studied and optimized, obtaining a limit of detection of 30U/mL CA125. The selectivity of the immunoassay was proven against different concentration solutions of Vascular Endothelial Growth Factor (VEGF) antigen as an unspecific protein when a blank signal was obtained for all tested solutions. Finally, preliminary experiments in human serum samples spiked with CA125 protein were also performed. Therefore, the proposed system could represent a powerful point-of-care tool for the next generation technology for detecting and monitoring cancer biomarkers at early stages by taking advantage of nowadays gadgets with enhanced features such as smartphones.