Foodborne illnesses are a growing concern for the food industry and consumers, with millions of cases reported every year. Consequently, there is a critical need to develop rapid, sensitive, and inexpensive techniques for pathogen detection in order to mitigate this problem. However, current pathogen detection strategies mainly include time-consuming laboratory methods and highly trained personnel. Electrochemical in-field biosensors offer a rapid, low-cost alternative to laboratory techniques, but the electrodes used in these biosensors require expensive nanomaterials to increase their sensitivity, such as noble metals (e.g., platinum, gold) or carbon nanomaterials (e.g., carbon nanotubes, or graphene). Herein, we report the fabrication of a highly sensitive and label-free laser-induced graphene (LIG) electrode that is subsequently functionalized with antibodies to electrochemically quantify the foodborne pathogen Salmonella enterica serovar Typhimurium. The LIG electrodes were produced by laser induction on polyimide film in ambient conditions, and hence circumvent the need for hightemperature, vacuum environment, and metal seed catalysts commonly associated with graphene-based electrodes fabricated via chemical vapor deposition processes. After functionalization with Salmonella-antibodies, the LIG biosensors were able to detect live Salmonella in chicken broth across a wide linear range (25 to 10 5 CFU mL -1 ) and with a low detection limit (13 ± 7 CFU mL -1 ; n = 3, mean ± standard deviation). These results were acquired with an average response time of 22 minutes without the need for sample preconcentration or redox labeling techniques. Moreover, these LIG immunosensors displayed high selectivity as demonstrated by non-significant response to other bacteria strains. These results demonstrate how LIG-based electrodes can be used for electrochemical immunosensing in general and, more specifically, could be used as a viable option for rapid, low-cost pathogen detection in food processing facilities before contaminated foods reach the consumer.
Carbon nanomaterials such as graphene exhibit unique material properties such as high electrical conductivity, surface area, and biocompatibility that have the potential to significantly improve the performance of electrochemical sensors. Since in-field electrochemical sensors are typically disposable, they require materials that are amenable to low-cost, high-throughput, and scalable manufacturing. Conventional graphene devices based on low-yield chemical vapor deposition techniques are too expensive for such applications, while low-cost alternatives such as screen and inkjet printing do not possess sufficient control over electrode geometry to achieve favorable electrochemical sensor performance. In this work, aerosol jet printing (AJP) is used to create high-resolution (~40 µm line width) interdigitated electrodes (IDEs) on flexible substrates, which are then converted into histamine sensors by covalently linking monoclonal antibodies to oxygen moieties created on the graphene surface through a CO2 thermal annealing process. The resulting electrochemical sensors exhibit a wide histamine sensing range of 6. 25-200 ppm (56.25 µM -1.8 mM) and a low detection limit of 3.41 ppm (30.7 µM) within actual tuna broth samples. These sensor metrics are significant since histamine levels over 50 ppm in fish induce adverse health effects including severe allergic reactions (e.g., Scombroid food poisoning).Beyond the histamine case study presented here, the AJP and functionalization process can likely be generalized to a diverse range of sensing applications including environmental toxin detection, foodborne pathogen detection, wearable health monitoring, and health diagnostics.
Cellulose nanofibers were extracted from sisal and incorporated at different concentrations (0-5%) into cassava starch to produce nanocomposites. Films' morphology, thickness, transparency, swelling degree in water, water vapor permeability (WVP) as well as thermal and mechanical properties were studied. Cellulose nanofiber addition affected neither thickness (56.637 6 2.939 mm) nor transparency (2.97 6 1.07 mm 21 ). WVP was reduced until a cellulose nanofiber content of 3.44%. Tensile force was increased up to a nanocellulose concentration of 3.25%. Elongation was decreased linearly upon cellulose nanofiber addition. Among all films, the greatest Young's modulus was 2.2 GPa. Cellulose nanofibers were found to reduce the onset temperature of thermal degradation, although melting temperature and enthalpy were higher for the nanocomposites. Because cellulose nanofibers were able to improve key properties of the films, the results obtained here can pave the route for the development and large-scale production of novel biodegradable packaging materials.
Rapid, inexpensive, and easy-to-use coronavirus disease 2019 (COVID-19) home tests are key tools in addition to vaccines in the world-wide fight to eliminate national and local shutdowns. However, currently available tests for SARS-CoV-2, the virus that causes COVID-19, are too expensive, painful, and irritating, or not sufficiently sensitive for routine, accurate home testing. Herein, we employ custom-formulated graphene inks and aerosol jet printing (AJP) to create a rapid electrochemical immunosensor for direct detection of SARS-CoV-2 Spike Receptor-Binding Domain (RBD) in saliva samples acquired non-invasively. This sensor demonstrated limits of detection that are considerably lower than most commercial SARS-CoV-2 antigen tests (22.91 ± 4.72 pg/mL for Spike RBD and 110.38 ± 9.00 pg/mL for Spike S1) as well as fast response time (~30 mins), which was facilitated by the functionalization of printed graphene electrodes in a single-step with SARS-CoV-2 polyclonal antibody through the carbodiimide reaction without the need for nanoparticle functionalization or secondary antibody or metallic nanoparticle labels. This immunosensor presents a wide linear sensing range from 1 to 1000 ng/mL and does not react with other coexisting influenza viruses such as H1N1 hemagglutinin. By combining high-yield graphene ink synthesis, automated printing, high antigen selectivity, and rapid testing capability, this work offers a promising alternative to current SARS-CoV-2 antigen tests.
Irrigation water is a primary source of fresh produce contamination by bacteria during the preharvest, particularly in hydroponic systems where the control of pests and pathogens is a major challenge. In this work, we demonstrate the development of a Listeria biosensor using platinum interdigitated microelectrodes (Pt-IME). The sensor is incorporated into a particle/sediment trap for the real-time analysis of irrigation water in a hydroponic lettuce system. We demonstrate the application of this system using a smartphone-based potentiostat for rapid on-site analysis of water quality. A detailed characterization of the electrochemical behavior was conducted in the presence/absence of DNA and Listeria spp., which was followed by calibration in various solutions with and without flow. In flow conditions (100 mL samples), the aptasensor had a sensitivity of 3.37 ± 0.21 kΩ log-CFU−1 mL, and the LOD was 48 ± 12 CFU mL−1 with a linear range of 102 to 104 CFU mL−1. In stagnant solution with no flow, the aptasensor performance was significantly improved in buffer, vegetable broth, and hydroponic media. Sensor hysteresis ranged from 2 to 16% after rinsing in a strong basic solution (direct reuse) and was insignificant after removing the aptamer via washing in Piranha solution (reuse after adsorption with fresh aptamer). This is the first demonstration of an aptasensor used to monitor microbial water quality for hydroponic lettuce in real time using a smartphone-based acquisition system for volumes that conform with the regulatory standards. The aptasensor demonstrated a recovery of 90% and may be reused a limited number of times with minor washing steps.
Foodborne pathogens are a major concern for public health. We demonstrate for the first time a partially automated sensing system for rapid (~17 min), label-free impedimetric detection of Escherichia coli spp. in food samples (vegetable broth) and hydroponic media (aeroponic lettuce system) based on temperature-responsive poly(N-isopropylacrylamide) (PNIPAAm) nanobrushes. This proof of concept (PoC) for the Sense-Analyze-Respond-Actuate (SARA) paradigm uses a biomimetic nanostructure that is analyzed and actuated with a smartphone. The bio-inspired soft material and sensing mechanism is inspired by binary symbiotic systems found in nature, where low concentrations of bacteria are captured from complex matrices by brush actuation driven by concentration gradients at the tissue surface. To mimic this natural actuation system, carbon-metal nanohybrid sensors were fabricated as the transducer layer, and coated with PNIPAAm nanobrushes. The most effective coating and actuation protocol for E. coli detection at various temperatures above/below the critical solution temperature of PNIPAAm was determined using a series of electrochemical experiments. After analyzing nanobrush actuation in stagnant media, we developed a flow through system using a series of pumps that are triggered by electrochemical events at the surface of the biosensor. SARA PoC may be viewed as a cyber-physical system that actuates nanomaterials using smartphone-based electroanalytical testing of samples. This study demonstrates thermal actuation of polymer nanobrushes to detect (sense) bacteria using a cyber-physical systems (CPS) approach. This PoC may catalyze the development of smart sensors capable of actuation at the nanoscale (stimulus-response polymer) and macroscale (non-microfluidic pumping).
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