Thermal Pyocyanin Sensor Based on Molecularly Imprinted Polymers for the Indirect Detection of Pseudomonas aeruginosa.

Pseudomonas aeruginosa is a ubiquitous multi-drug-resistant bacterium, capable of causing serious illnesses and infections. This research focuses on the development of a thermal sensor for the indirect detection of P. aeruginosa infection using molecularly imprinted polymers (MIPs). This was achieved by developing MIPs for the detection of pyocyanin, the main toxin secreted by P. aeruginosa. To this end, phenazine was used as a dummy template, evaluating several polymeric compositions to achieve a selective MIP for pyocyanin recognition. The sensitivity of the synthesized MIPs was investigated by UV-vis analysis, with the best composition having a maximum rebinding capacity of 30 µmol g-1 and an imprinting factor (IF) of 1.59. Subsequently, the MIP particles were immobilized onto planar aluminum chips using an adhesive layer, to perform thermal resistance measurements at clinically relevant concentrations of pyocyanin (1.4-9.8 µM), achieving a limit of detection (LoD) of 0.347 ± 0.027 µM. The selectivity of the sensor was also scrutinized by subjecting the receptor to potential interferents. Furthermore, the rebinding was demonstrated in King's A medium, highlighting the potential of the sensor for the indirect detection of P. aeruginosa in complex fluids. The research culminates in the demonstration of the MIP-based sensor's applicability for clinical diagnosis. To achieve this goal, an experiment was performed in which the sensor was exposed to pyocyanin-spiked saliva samples, achieving a limit of detection of 0.569 ± 0.063 µM and demonstrating that this technology is suitable to detect the presence of the toxin even at the very first stage of its production.

Functionalized screen-printed electrodes for the thermal detection of Escherichia coli in dairy products.

Accurate and fast on-site detection of harmful microorganisms in food products is a key preventive step to avoid food-borne illness and product recall. In this study, screen-printed electrodes (SPEs) were functionalized via a facile strategy with surface imprinted polymers (SIPs). The SIP-coated SPEs were used in combination with the heat transfer method (HTM) for the real-time detection of Escherichia coli. The sensor was tested in buffer, with a reproducible and sensitive response that attained a limit of detection of 180 CFU/mL. Furthermore, selectivity was assessed by analyzing the sensor's response to C. sakazakii, K. pneumoniae and S. aureus as analogue strains. Finally, the device was successfully used for the detection of E. coli in spiked milk as proof-of-application, requiring no additional sample preparation. These results suggest the proposed thermal biosensor possesses the potential of becoming a tool for routine, on-site monitoring of E. coli in food safety applications.

Imprinted Polydimethylsiloxane-Graphene Oxide Composite Receptor for the Biomimetic Thermal Sensing of Escherichia coli.

This work presents an imprinted polymer-based thermal biomimetic sensor for the detection of Escherichia coli. A novel and facile bacteria imprinting protocol for polydimethylsiloxane (PDMS) films was investigated, and these receptor layers were functionalized with graphene oxide (GO) in order to improve the overall sensitivity of the sensor. Upon the recognition and binding of the target to the densely imprinted polymers, a concentration-dependent measurable change in temperature was observed. The limit of detection attained for the sensor employing PDMS-GO imprints was 80 ± 10 CFU/mL, a full order lower than neat PDMS imprints (670 ± 140 CFU/mL), illustrating the beneficial effect of the dopant on the thermo-dynamical properties of the interfacial layer. A parallel benchmarking of the thermal sensor with a commercial impedance analyzer was performed in order to prove the possibility of using the developed PDMS-GO receptors with multiple readout platforms. Moreover, S. aureus, C. sakazakii and an additional E. coli strain were employed as analogue species for the assessment of the selectivity of the device. Finally, because of the potential that this biomimetic platform possesses as a low-cost, rapid, and on-site tool for monitoring E. coli contamination in food safety applications, spiked fruit juice was analyzed as a real sample. Reproducible and sensitive results fulfill the limit requirements of the applicable European microbiological regulation.

Thermal Detection of Glucose in Urine Using a Molecularly Imprinted Polymer as a Recognition Element.

Glucose bio-sensing technologies have received increasing attention in the last few decades, primarily due to the fundamental role that glucose metabolism plays in diseases (e.g., diabetes). Molecularly imprinted polymers (MIPs) could offer an alternative means of analysis to a field that is traditionally dominated by enzyme-based devices, posing superior chemical stability, cost-effectiveness, and ease of fabrication. Their integration into sensing devices as recognition elements has been extensively studied with different readout methods such as quartz-crystal microbalance or impedance spectroscopy. In this work, a dummy imprinting approach is introduced, describing the synthesis and optimization of a MIP toward the sensing of glucose. Integration of this polymer into a thermally conductive receptor layer was achieved by micro-contact deposition. In essence, the MIP particles are pressed into a polyvinyl chloride adhesive layer using a polydimethylsiloxane stamp. The prepared layer is then evaluated with the so-called heat-transfer method, allowing the determination of the specificity and the sensitivity of the receptor layer. Furthermore, the selectivity was assessed by analyzing the thermal response after infusion with increasing concentrations of different saccharide analogues in phosphate-buffered saline (PBS). The obtained results show a linear range of the sensor of 0.0194-0.3300 mM for the detection of glucose in PBS. Finally, a potential application of the sensor was demonstrated by exposing the receptor layer to increasing concentrations of glucose in human urine samples, demonstrating a linear range of 0.0444-0.3300 mM. The results obtained in this paper highlight the applicability of the sensor both in terms of non-invasive glucose monitoring and for the analysis of food samples.

Imprinted Polymers as Synthetic Receptors in Sensors for Food Safety.

Foodborne illnesses represent high costs worldwide in terms of medical care and productivity. To ensure safety along the food chain, technologies that help to monitor and improve food preservation have emerged in a multidisciplinary context. These technologies focus on the detection and/or removal of either biological (e.g., bacteria, virus, etc.) or chemical (e.g., drugs and pesticides) safety hazards. Imprinted polymers are synthetic receptors able of recognizing both chemical and biological contaminants. While numerous reviews have focused on the use of these robust materials in extraction and separation applications, little bibliography summarizes the research that has been performed on their coupling to sensing platforms for food safety. The aim of this work is therefore to fill this gap and highlight the multidisciplinary aspects involved in the application of imprinting technology in the whole value chain ranging from IP preparation to integrated sensor systems for the specific recognition and quantification of chemical and microbiological contaminants in food samples.

Modular Science Kit as a support platform for STEM learning in primary and secondary school.

The need to develop interest in STEM (science, technology, engineering, and mathematics) skills in young pupils has driven many educational systems to include STEM as a subject in primary schools. In this work, a science kit aimed at children from 8 to 14 years old is presented as a support platform for an innovative and stimulating approach to STEM learning. The peculiar design of the kit, based on modular components, is aimed to help develop a multitude of skills in the young students, dividing the learning process into two phases. During phase 1 the pupils build the experimental setup and visualize the scientific phenomena, while in phase 2, they are introduced and challenged to understand the principles on which these phenomena are based, guided by a handbook. This approach aims at making the experience more inclusive, stimulating the interest and passion of the pupils for scientific subjects.

A Molecularly Imprinted Polymer-based Dye Displacement Assay for the Rapid Visual Detection of Amphetamine in Urine.

The rapid sensing of drug compounds has traditionally relied on antibodies, enzymes and electrochemical reactions. These technologies can frequently produce false positives/negatives and require specific conditions to operate. Akin to antibodies, molecularly imprinted polymers (MIPs) are a more robust synthetic alternative with the ability to bind a target molecule with an affinity comparable to that of its natural counterparts. With this in mind, the research presented in this article introduces a facile MIP-based dye displacement assay for the detection of (±) amphetamine in urine. The selective nature of MIPs coupled with a displaceable dye enables the resulting low-cost assay to rapidly produce a clear visual confirmation of a target's presence, offering huge commercial potential. The following manuscript characterizes the proposed assay, drawing attention to various facets of the sensor design and optimization. To this end, synthesis of a MIP tailored towards amphetamine is described, scrutinizing the composition and selectivity (ibuprofen, naproxen, 2-methoxphenidine, quetiapine) of the reported synthetic receptor. Dye selection for the development of the displacement assay follows, proceeded by optimization of the displacement process by investigating the time taken and the amount of MIP powder required for optimum displacement. An optimized dose-response curve is then presented, introducing (±) amphetamine hydrochloride (0.01-1 mg mL-1) to the engineered sensor and determining the limit of detection (LoD). The research culminates in the assay being used for the analysis of spiked urine samples (amphetamine, ibuprofen, naproxen, 2-methoxphenidine, quetiapine, bupropion, pheniramine, bromopheniramine) and evaluating its potential as a low-cost, rapid and selective method of analysis.

Rapid Colorimetric Screening of Elevated Phosphate in Urine: A Charge-Transfer Interaction.

A charge-transfer (CT) interaction between 1,3,5-trinitro-2,4-dimethylbenzene (TNX) and anionic phosphate is evaluated, yielding a high band electronic transfer interaction that can be observed as a distinct color change when phosphate is present in solution. The induced interaction was studied using 1H NMR, UV-visible, and Fourier transform infrared spectroscopies. The stoichiometric determination of the interaction was divined by means of continuous variation, applying the Schaeppi-Treadwell method to calculate the binding constant (k). Furthermore, the effect of the polarity of solvents toward the generation of the CT interaction was examined, with multiple solvents considered. Complex deconstruction studies were undertaken, examining the effects of water on complex destruction and understanding the volumes needed to hinder the CT interaction potency. Specificity and selectivity of the CT interaction were also studied against other biologically relevant species (CH3CH2OH, Na+, K+, Ca2+, Cl-, HCO3 -, F-, CH3COO-, and SO4 2-), assessing the capabilities of the assay to differentiate anionic species and counter cations that could act as interferences. The role of TNX concentration in CT formation was also analyzed, aiming to optimize the phosphate-sensing assay and improve its limit of detection. The sensing platform was subsequently used to study phosphate concentrations in urine samples to further understand its potential application in biomedical research. To validate the developed technique, urine samples were analyzed for their phosphate content with both the developed sensor and a validated vanadate-molybdate reagent. The results indicate that the sensing method is capable of accurately reporting elevated phosphate levels in urine samples in a rapid and sensitive manner, illustrating that the colorimetric test could be used as a prescreening test for conditions such as hyperphosphatemia or chronic kidney disease.