Staphylococcus aureus - Review on potential targets for sensors development.

Staphylococcus aureus (S. aureus) is accountable for a wide variety of clinical disease with a high rate of morbidity and mortality around the globe. It has a leading place into the ESKAPE group that includes six pathogens and exhibit multidrug resistance and are the major cause of healthcare associated infections: Enterococcus faecium, S. aureus, Klebsiella pneumoniae, Acinetobacter baumannii, Pseudomonas aeruginosa, and Enterobacter spp. A critical overview regarding the development of sensors for both S. aureus and his, more dangerous alter ego, Methicillin-resistant S. aureus (MRSA) was presented focusing on the bacteria targets starting with the detection of the whole cell, up to specific wall components, toxins or other virulence factors. The literature data was systematically assessed having in sight the design of the sensing platforms, the analytical performances, and possible courses of action to be implemented in real practice as point-of-care (POC) devices. Moreover, a distinct section was dedicated to commercially available devices and out of the box approaches, namely the use of bacteriophages as an alternative to antimicrobial therapy and as sensors modifiers. The reviewed sensors and devices were discussed in terms of their suitability for different biosensing applications, in early screening of contamination regarding food analysis, environmental monitoring and in clinical diagnosis.

Proof of Concept for the Detection with Custom Printed Electrodes of Enterobactin as a Marker of Escherichia coli.

The rapid and decentralized detection of bacteria from biomedical, environmental, and food samples has the capacity to improve the conventional protocols and to change a predictable outcome. Identifying new markers and analysis methods represents an attractive strategy for the indirect but simpler and safer detection of pathogens that could replace existing methods. Enterobactin (Ent), a siderophore produced by Escherichia coli or other Gram-negative bacteria, was studied on different electrode materials to reveal its electrochemical fingerprint-very useful information towards the detection of the bacteria based on this analyte. The molecule was successfully identified in culture media samples and a future goal is the development of a rapid antibiogram. The presence of Ent was also assessed in wastewater and treated water samples collected from the municipal sewage treatment plant, groundwater, and tap water. Moreover, a custom configuration printed on a medical glove was employed to detect the target in the presence of another bacterial marker, namely pyocyanin (PyoC), that being a metabolite specific of another pathogen bacterium, namely Pseudomonas aeruginosa. Such new mobile and wearable platforms offer considerable promise for rapid low-cost on-site screening of bacterial contamination.

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.

Electrochemical Fingerprints of Illicit Drugs on Graphene and Multi-Walled Carbon Nanotubes.

Illicit drugs use and abuse 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 increased tremendously in recent years. Nanomaterials have gained much attention over the last decade in the development of sensors for a myriad of applications. The applicability of these nanomaterials, functionalized or not, significantly increases and it is therefore highly suitable for use in the detection of illicit drugs. We have assessed the suitability of various nanoplatforms, such as graphene (GPH), multi-walled carbon nanotubes (MWCNTs), gold nanoparticles (AuNPs) and platinum nanoparticles (PtNPs) for the electrochemical detection of illicit drugs. GPH and MWCNTs were chosen as the most suitable platforms and cocaine, 3,4-methylendioxymethamfetamine (MDMA), 3-methylmethcathinone (MMC) and α-pyrrolidinovalerophenone (PVP) were tested. Due to the hydrophobicity of the nanomaterials-based platforms which led to low signals, two strategies were followed namely, pretreatment of the electrodes in sulfuric acid by cyclic voltammetry and addition of Tween 20 to the detection buffer. Both strategies led to an increase in the oxidation signal of illicit drugs. Binary mixtures of illicit drugs with common adulterants found in street samples were also investigated. The proposed strategies allowed the sensitive detection of illicit drugs in the presence of most adulterants. The suitability of the proposed sensors for the detection of illicit drugs in spiked wastewaters was finally assessed.

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.

An overview of the detection of serotonin and dopamine with graphene-based sensors.

The development of rapid and sensitive devices for the simultaneous detection of neurotransmitters has critical implications for the clinical field and for the management of several diseases. Parkinson's, Alzheimer's disease, autism, schizophrenia, depression and anxiety are major healthcare challenges for which early diagnostics and personalized therapy are of great concern. Carbon-based nanomaterials and especially graphene-based nanomaterials associated with different architectures have been extensively studied and continue to represent the first line of approach in the development of nanoplatforms for electrochemical sensors. The simultaneous detection of analytes represents a critical point that could be addressed by designing new materials with the capacity to resolve their electrochemical signals. The results can be presented as a matrix that offers a broader viewpoint toward the balance of the neurotransmitter levels that are correlated with clinical symptoms for personalized diagnosis. The goal is to describe and evaluate, in a critical manner, the elaboration of graphene-based sensors that can be included in clinical applications. A major check point discussed throughout the paper is represented by the interference between different neurotransmitters that appear due to their overlapping signals and the strategies to address them to achieve simultaneous detection of as many molecules as possible and to be similar to in vivo experiments.

Electrochemical Non-Enzymatic Detection of Glucose Based on 3D Electroformed Copper on Ni Foam Nanostructures.

Despite the fact that the electrochemical biosensors based on glucose oxidase represent the golden standard for the management of diabetes, the elaboration of nonenzymatic sensors became extensively studied as an out-of-the-box concept that aims to simplify the existing approach. An important point of view is represented by the low price of the sensing device that has positive effects for both end-users and healthcare systems. The enzyme-free sensors based on low-cost materials such as transition metals have similar analytical properties to the commercial ones while eliminating the issues associated with the presence of the enzyme, such as the stability issues and limited shelf-life. The development of nanoporous nanomaterials for biomedical applications and electrocatalysis was referred to as an alternative to the conventional methods due to their enlarged area, electrical properties, ease of functionalization and not least to their low cost. Herein, we report the development of an electrochemical nonenzymatic sensor for glucose based on 3D copper nanostructures with Ni foams as promotor of the enhanced nanoporous morphology. The sensors were successfully tested in the presence of the designated target, even in the presence of common interference agents found in biological samples.

Selective Detection of Folic Acid Using 3D Polymeric Structures of 3-Carboxylic Polypyrrole.

The detection of folic acid in biological samples or pharmaceutical products is of great importance due to its implications in the biological functions of the human body, along with the development and growth of the fetus. The deficiency of folic acid can be reversed by the intake of different pharmaceutical formulations or alimentary products fortified with this molecule. The elaboration of sensing platforms represents a continuous work in progress, a task in which the use of conductive polymers modified with different functionalities represents one of the outcoming strategies. The possibility of manipulating their morphology with the use of templates or surfactants represents another advantage. A sensing platform based on carboxylic functionalized polypyrrole was synthesized via the electrochemical approach in the presence of a polymeric surfactant on a graphite-based surface. The sensor was able to detect the folic acid from 2.5 µM to 200 µM with a calculated limited of detection of 0.8 µM. It was employed for the detection of the analyte from commercial human serum and pharmaceutical products with excellent recovery rates. The results were double checked using an optimized spectrophotometric procedure that confirmed furthermore the performances of the sensor related to real samples assessment.

Synergic action of thermosensitive hydrogel and Au/Ag nanoalloy for sensitive and selective detection of pyocyanin.

The rapid detection of bacterial strains has become a major topic thoroughly discussed across the biomedical field. Paired with the existence of nosocomial pathogen agents that imply extreme medical and financial challenges throughout diagnosis and treatment, the development of rapid and easy-to-use sensing devices has gained an increased amount of attention. Moreover, antibiotic resistance considered by World Health Organization as one of the "biggest threats to global health, food security, and development today" enables this topic as high priority. Pseudomonas aeruginosa, one of the most ubiquitous bacterial strains, has various quorum-sensing systems that are a direct cause of their virulence. One of them is represented by pyocyanin, a blue pigment with electroactive properties that is synthesized from early stages of bacterial colonization. Thus, the sensitive detection of this biomarker could enable a personalized and efficient therapy. It was achieved with the development of an electrochemical sensor based on a thermosensitive polymer, modified with Au/Ag nanoalloy for the rapid and accurate detection of pyocyanin, a virulence biomarker of Pseudomonas aeruginosa. The sensor displayed a linear range from 0.12 to 25 µM, and a limit of detection of 0.04 µM (signal/noise = 3). It was successfully tested in real samples spiked with the target analyte without any pretreatment other than a dilution step. The detection of pyocyanin with high recovery in whole blood in a time frame of 5-10 min from the moment of collection was performed with this electrochemical sensor. Graphical abstract.

A Nanocomposite Based on Reduced Graphene and Gold Nanoparticles for Highly Sensitive Electrochemical Detection of Pseudomonas aeruginosa through Its Virulence Factors.

Pyoverdine is a fluorescent siderophore produced by Pseudomonas aeruginosa that can be considered as a detectable marker in nosocomial infections. The presence of pyoverdine in water can be directly linked to the presence of the P. aeruginosa, thus being a nontoxic and low-cost marker for the detection of biological contamination. A novel platform was developed and applied for the electrochemical selective and sensitive detection of pyoverdine, based on a graphene/graphite-modified screen-printed electrode (SPE) that was electrochemically reduced and decorated with gold nanoparticles (NPs). The optimized sensor presenting higher sensitivity towards pyoverdine was successfully applied for its detection in real samples (serum, saliva, and tap water), in the presence of various interfering species. The excellent analytical performances underline the premises for an early diagnosis kit of bacterial infections based on electrochemical sensors.

Finger-Based Printed Sensors Integrated on a Glove for On-Site Screening Of Pseudomonas aeruginosa Virulence Factors.

Early screening of clinically relevant pathogens in the environment is a highly desirable goal in clinical care, providing precious information that will improve patient-care outcomes. In this work, a glove-based electrochemical sensor has been designed for point-of-use screening of Pseudomonas aeruginosa's virulence factors. The methodology used for the elaboration of the fabric platform relied on printing the conductive inks on the index and middle fingers of the glove, with the goal of screening pyocyanin and pyoverdine targets. The analytical signatures of the analytes were recorded in about 4 min, via the rapid and selective square-wave-voltammetry technique. Finger-based sensors display good performance and discrimination between the targets and potential interferences, along with good reproducibility. The sensors featured linearity over the 0.01-0.1 µM range for pyocyanin and 5-50 µM range for pyoverdine, with sensitivities of 2.51 µA/µM for pyocyanin and 1.09 nA/µM for pyoverdine ( R2 = 0.990 and 0.995, respectively) and detection limits of 3.33 nM for pyocyanin and 1.66 µM for pyoverdine. Moreover, the sensors were tested in binary mixtures of analytes, with successful outcomes. In order to gain information from the surrounding environment, the active electronic areas of the printed fingers were coated with a conductive hydrogel matrix, and relevant target surfaces were "swiped for notification" of contaminants. The simple fabrication, low-cost, and reusability of the proposed glove are likely to underpin the progressive drive of wearable sensors toward decentralized environmental and healthcare applications.

Sensitive detection of pyoverdine with an electrochemical sensor based on electrochemically generated graphene functionalized with gold nanoparticles.

The design and development of an electrochemical sensor for the sensitive and selective determination of pyoverdine, a virulence factor secreted by Pseudomonas aeruginosa, bacteria involved in nosocomial infections is presented in this work. The presence of pyoverdine in water and body fluids samples can be directly linked to the presence of the Pseudomonas bacteria, thus being a nontoxic and low cost marker for the detection of water pollution as well as for the biological contamination of other media. The sensor was elaborated using layer-by-layer technique for the deposition of a graphene­gold nanoparticles composite film on the graphite-based screen printed electrode, from aqueous suspension. Under optimal conditions, the electrochemical signal corresponding to the pyoverdine oxidation process was proportional to its concentration, showing a wide linear range from 1 to 100µmolL-1 and a detection limit of 0.33µmolL-1. This sensor discriminate with satisfactory recoveries the target analyte in different real matrices and also exhibited low response to other interfering species, proving that this technique is promising for medical and environmental applications. In addition, the proposed nanocomposite platform presented good reproducibility, high and long term stability, the sensitivity for pyoverdine remain unchanged after being stored at 4°C for four weeks.

Detection of Dopamine by a Biomimetic Electrochemical Sensor Based on Polythioaniline-Bridged Gold Nanoparticles.

A new biomimetic electrochemical sensor was developed for the detection of dopamine based on a glassy carbon electrode modified with electrochemically generated gold nanoparticles. The preparation of the polymer is simple and cost-effective, achieving the polymerization of thioaniline and generation of gold nanoparticles in a single step by cyclic voltammetry, in the presence of the target molecule, dopamine. After extraction, the imprinted polymer exhibits high sensitivity and selectivity for dopamine. Moreover, the developed imprinted polymer film allows the fast, direct detection of dopamine without the need of a redox mediator. The formation of a self-assembled monolayer of the monomer prior to electropolymerization ensures the adherence of the film onto the electrode surface, conferring good stability to the sensor (over two weeks). Cyclic voltammetry, electrochemical impedance spectroscopy, atomic force microscopy, scanning electron microscopy, and energy dispersive X-ray spectroscopy were used for the complete characterization of the developed sensor, and differential pulse voltammetry was used for its testing.

Surface Modeling of Nanopatterned Polymer Films Obtained by Colloidal Templated Electropolymerization.

Nanostructured polypyrrole surfaces are fabricated by a one step colloidal templating approach, involving simultaneous polystyrene bead deposition with the electropolymerization of the pyrrole monomer. Using response surface modeling, the influence of several experimental parameters was studied, following as response the resulted film's thickness and the nanopattern's surface density. Polystyrene beads of 100 nm were immobilized by the electropolymerization of pyrrole and the spheres were dissolved in tetrahydrofuran at room temperature. The obtained nanostructures were analyzed by atomic force microscopy (AFM) and the results served to build the mathematical model (central composite face centered design, quadratic model) best describing the correlations between variables and responses. The obtained response surface model revealed the specific influence of each of the studied variables over the followed responses and it also allowed the controlled fabrication of nanopatterned surfaces.

Electrochemical sensors based on carbon nanomaterials for acetaminophen detection: A review.

This study describes the advancements made over the last five years in the development of electrochemical sensors and biosensors for acetaminophen detection. This study reviews the different configurations based on unmodified and chemically modified carbon nanotubes and graphene. The influence of various modifiers on the two types of materials is presented along with their role on the enhancement of the selectivity and sensitivity of (bio)sensors. The review is focused on a comparative description of the applications of carbon-based nanomaterials towards acetaminophen detection and presents the results in a critical manner.

Micro- to nanostructured poly(pyrrole-nitrilotriacetic acid) films via nanosphere templates: applications to 3D enzyme attachment by affinity interactions.

We report the combination of latex nanosphere lithography with electropolymerization of N-substituted pyrrole monomer bearing a nitrilotriacetic acid (NTA) moiety for the template-assisted nanostructuration of poly(pyrrole-NTA) films and their application for biomolecule immobilization. The electrodes were modified by casting latex beads (100 or 900 nm in diameter) on their surface followed by electropolymerization of the pyrrole-NTA monomer and the subsequent chelation of Cu(2+) ions. The dissolution of the nanobeads leads then to a nanostructured polymer film with increased surface. Thanks to the versatile affinity interactions between the (NTA)Cu(2+) complex and histidine- or biotin-tagged proteins, both tyrosinase and glucose oxidase were immobilized on the modified electrode. Nanostructuration of the polypyrrole via nanosphere lithography (NSL) using 900- and 100-nm latex beads allows an increase in surface concentration of enzymes anchored on the functionalized polypyrrole electrode. The nanostructured enzyme electrodes were characterized by fluorescence microscopy, 3D laser scanning confocal microscopy, and scanning electron microscopy. Electrochemical studies demonstrate the increase in the amount of immobilized biomolecules and associated biosensor performances when achieving NSL compared to conventional polymer formation without bead template. In addition, the decrease in nanobead diameter from 900 to 100 nm provides an enhancement in biosensor performance. Between biosensors based on films polymerized without nanobeads and with 100-nm nanobeads, maximum current density values increase from 4 to 56 µA cm(-2) and from 7 to 45 µA cm(-2) for biosensors based on tyrosinase and glucose oxidase, respectively.