Planar Interdigitated Aptasensor for Flow-Through Detection of Listeria spp. in Hydroponic Lettuce Growth Media

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.

[1]  S. Wilks,et al.  The survival of Escherichia coli O157 on a range of metal surfaces. , 2005, International journal of food microbiology.

[2]  Eric S. McLamore,et al.  Self-referencing optrodes for measuring spatially resolved, real-time metabolic oxygen flux in plant systems , 2010, Planta.

[3]  I. Peiris Listeria monocytogenes, a Food-Borne Pathogen , 1991, Microbiological reviews.

[4]  Michael J. Schöning,et al.  Study of Interdigitated Electrode Arrays Using Experiments and Finite Element Models for the Evaluation of Sterilization Processes , 2015, Sensors.

[5]  I. Suni,et al.  Detection of Listeria Monocytogenes by Electrochemical Impedance Spectroscopy , 2013 .

[6]  Yue Rong,et al.  SNAPS: Sensor Analytics Point Solutions for Detection and Decision Support Systems , 2019, Sensors.

[7]  Catherine Disselhorst-Klug,et al.  Simulation analysis of the ability of different types of multi-electrodes to increase selectivity of detection and to reduce cross-talk. , 2003, Journal of electromyography and kinesiology : official journal of the International Society of Electrophysiological Kinesiology.

[8]  Susan S. Ellenberg Food and Drug Administration (FDA) , 2005 .

[9]  Klaus D Goepel,et al.  Implementation of an Online Software Tool for the Analytic Hierarchy Process (AHP-OS) , 2018, International Journal of the Analytic Hierarchy Process.

[10]  E. Alocilja,et al.  Sensor-as-a-Service: Convergence of Sensor Analytic Point Solutions (SNAPS) and Pay-A-Penny-Per-Use (PAPPU) Paradigm as a Catalyst for Democratization of Healthcare in Underserved Communities , 2020, Diagnostics.

[11]  Eric S McLamore,et al.  Actuation of chitosan-aptamer nanobrush borders for pathogen sensing. , 2018, The Analyst.

[12]  Soojin Jun,et al.  ABE-Stat, a Fully Open-Source and Versatile Wireless Potentiostat Project Including Electrochemical Impedance Spectroscopy , 2019, Journal of the Electrochemical Society.

[13]  S. Flint,et al.  Colonisation of lettuce by Listeria Monocytogenes , 2018, International Journal of Food Science & Technology.

[14]  Chii-Wann Lin,et al.  Advances in rapid detection methods for foodborne pathogens. , 2014, Journal of microbiology and biotechnology.

[15]  S. Stroika,et al.  Multistate Outbreak of Listeriosis Associated with Packaged Leafy Green Salads, United States and Canada, 2015–2016 , 2019, Emerging infectious diseases.

[16]  D. Nyachuba Foodborne illness: is it on the rise? , 2010, Nutrition reviews.

[17]  César Fernández-Sánchez,et al.  Reusable conductimetric array of interdigitated microelectrodes for the readout of low-density microarrays. , 2014, Analytica chimica acta.

[18]  V. Wu,et al.  Gold Nanoparticle-Modified Carbon Electrode Biosensor for the Detection of Listeria monocytogenes , 2013 .

[19]  A. Bhunia,et al.  Antibody–aptamer functionalized fibre‐optic biosensor for specific detection of Listeria monocytogenes from food , 2010, Journal of applied microbiology.

[20]  Arun K. Bhunia,et al.  Detection of Low Levels of Listeria monocytogenes Cells by Using a Fiber-Optic Immunosensor , 2004, Applied and Environmental Microbiology.

[21]  Yanbin Li,et al.  A label-free, microfluidics and interdigitated array microelectrode-based impedance biosensor in combination with nanoparticles immunoseparation for detection of Escherichia coli O157:H7 in food samples , 2007 .

[22]  D. Vanegas,et al.  A paper based graphene-nanocauliflower hybrid composite for point of care biosensing , 2016, SPIE Commercial + Scientific Sensing and Imaging.

[23]  L. Warne,et al.  Electrophysics of micromechanical comb actuators , 1995 .

[24]  E. Alocilja,et al.  A high density microelectrode array biosensor for detection of E. coli O157:H7. , 2005, Biosensors & bioelectronics.

[25]  Ronghui Wang,et al.  TiO2 nanowire bundle microelectrode based impedance immunosensor for rapid and sensitive detection of Listeria monocytogenes. , 2009, Nano letters.

[26]  Ji-Young Ahn,et al.  Analytical bioconjugates, aptamers, enable specific quantitative detection of Listeria monocytogenes. , 2015, Biosensors & bioelectronics.

[27]  Steve Tung,et al.  Efficient separation and sensitive detection of Listeria monocytogenes using an impedance immunosensor based on magnetic nanoparticles, a microfluidic chip, and an interdigitated microelectrode. , 2012, Journal of food protection.

[28]  V. Gold Compendium of chemical terminology , 1987 .

[29]  Lin Yang,et al.  Impedance DNA Biosensor Using Electropolymerized Polypyrrole/Multiwalled Carbon Nanotubes Modified Electrode , 2006 .

[30]  Maria C. DeRosa,et al.  Challenges and Opportunities for Small Molecule Aptamer Development , 2012, Journal of nucleic acids.

[31]  Ren Yi,et al.  Electronic nose classification and differentiation of bacterial foodborne pathogens based on support vector machine optimized with particle swarm optimization algorithm , 2019, Journal of Food Process Engineering.

[32]  Gunawan Witjaksono bin Djaswadi,et al.  Simulation of Interdigitated Electrodes (IDEs) Geometry Using COMSOL Multiphysics , 2018, 2018 International Conference on Intelligent and Advanced System (ICIAS).

[33]  M. Wiedmann,et al.  Diversity of Listeria Species in Urban and Natural Environments , 2012, Applied and Environmental Microbiology.

[34]  Eric S. McLamore,et al.  Impedance biosensor for the rapid detection of Listeria spp. based on aptamer functionalized Pt-interdigitated microelectrodes array , 2016, SPIE Commercial + Scientific Sensing and Imaging.

[35]  Eric S McLamore,et al.  A comparative study of carbon-platinum hybrid nanostructure architecture for amperometric biosensing. , 2014, The Analyst.

[36]  R. Bashir,et al.  Dielectrophoretic separation and manipulation of live and heat-treated cells of Listeria on microfabricated devices with interdigitated electrodes , 2002 .

[37]  P. Chaturvedi,et al.  A self-referencing biosensor for real-time monitoring of physiological ATP transport in plant systems. , 2015, Biosensors & bioelectronics.

[38]  E. H. Garrett,et al.  Outbreaks Associated with Fresh Produce: Incidence, Growth, and Survival of Pathogens in Fresh and Fresh‐Cut Produce , 2001 .

[39]  G. Grüner,et al.  Charge transfer and charge transport on the double helix , 2003, cond-mat/0309360.

[40]  Li Mu,et al.  ssDNA aptamer-based column for simultaneous removal of nanogram per liter level of illicit and analgesic pharmaceuticals in drinking water. , 2011, Environmental science & technology.

[41]  S. Flint,et al.  Rapid attachment of Listeria monocytogenes to hydroponic and soil grown lettuce leaves , 2019, Food Control.

[42]  Mohammed Zourob,et al.  Rapid colorimetric sensing platform for the detection of Listeria monocytogenes foodborne pathogen. , 2016, Biosensors & bioelectronics.

[43]  L. Jacxsens,et al.  Microbial hazards in irrigation water: standards, norms, and testing to manage use of water in fresh produce primary production. , 2015 .

[44]  R. Moreira,et al.  Growth of Listeria monocytogenes and Listeria innocua on fresh baby spinach leaves: Effect of storage temperature and natural microflora , 2015 .

[45]  Edward M. Fox,et al.  Control of Listeria species food safety at a poultry food production facility. , 2015, Food microbiology.

[46]  Antje J. Baeumner,et al.  Characterization and Optimization of Interdigitated Ultramicroelectrode Arrays as Electrochemical Biosensor Transducers , 2004 .

[47]  Wei H Lai,et al.  Efficient separation and quantitative detection of Listeria monocytogenes based on screen-printed interdigitated electrode, urease and magnetic nanoparticles , 2017 .

[48]  Mieke Uyttendaele,et al.  Quantitative study of cross-contamination with Escherichia coli, E. coli O157, MS2 phage and murine norovirus in a simulated fresh-cut lettuce wash process , 2014 .

[49]  Eric S. McLamore,et al.  Food Processing and Waste Within the Nexus Framework , 2017 .

[50]  E. Migliorini,et al.  Practical guide to characterize biomolecule adsorption on solid surfaces (Review). , 2018, Biointerphases.

[51]  T. Livache,et al.  Early detection of bacteria using SPR imaging and event counting: experiments with Listeria monocytogenes and Listeria innocua , 2019, RSC advances.

[52]  Eric S McLamore,et al.  Laser Scribed Graphene Biosensor for Detection of Biogenic Amines in Food Samples Using Locally Sourced Materials , 2018, Biosensors.

[53]  V. Gold,et al.  Compendium of chemical terminology : IUPAC recommendations , 1987 .

[54]  K. Gibson,et al.  Risk of Human Pathogen Internalization in Leafy Vegetables During Lab-Scale Hydroponic Cultivation , 2019, Horticulturae.

[55]  R. Mutharasan,et al.  Rapid and sensitive immunodetection of Listeria monocytogenes in milk using a novel piezoelectric cantilever sensor. , 2013, Biosensors & bioelectronics.

[56]  Thomas Thundat,et al.  Impedimetric detection of pathogenic Gram-positive bacteria using an antimicrobial peptide from class IIa bacteriocins. , 2014, Analytical chemistry.

[57]  Hee-Jung Lee,et al.  An ELISA-on-a-Chip Biosensor System for Early Screening of Listeria monocytogenes in Contaminated Food Products , 2009 .

[58]  Mahmoud Almasri,et al.  A micromachined impedance biosensor for accurate and rapid detection of E. coli O157:H7 , 2013 .

[59]  Ailiang Chen,et al.  Replacing antibodies with aptamers in lateral flow immunoassay. , 2015, Biosensors & bioelectronics.

[60]  A. B. Marks The Risks We Are Willing to Eat: Food Imports and Safety , 2015 .

[61]  Yanbin Li,et al.  Interdigitated array microelectrode based impedance biosensor coupled with magnetic nanoparticle-antibody conjugates for detection of Escherichia coli O157:H7 in food samples. , 2007, Biosensors & bioelectronics.

[62]  M. Schöning,et al.  Planar and 3D interdigitated electrodes for biosensing applications: The impact of a dielectric barrier on the sensor properties , 2014 .

[63]  Reversible Regulation of Aptamer Activity with Effector-Responsive Hairpin Oligonucleotides , 2013, Journal of laboratory automation.

[64]  Y. Massoud,et al.  Modeling and analysis of differential signaling for minimizing inductive crosstalk , 2001, Proceedings of the 38th Design Automation Conference (IEEE Cat. No.01CH37232).

[65]  Minhaz Uddin Ahmed,et al.  A bacteriophage endolysin-based electrochemical impedance biosensor for the rapid detection of Listeria cells. , 2012, The Analyst.

[66]  Yang Zhang,et al.  Recent Advances in Aptamer Discovery and Applications , 2019, Molecules.

[67]  A. Bhunia,et al.  Microscale electronic detection of bacterial metabolism , 2002 .

[68]  Y Rong,et al.  Post hoc support vector machine learning for impedimetric biosensors based on weak protein-ligand interactions. , 2018, The Analyst.

[69]  Zhixian Gao,et al.  Determination of Listeria Monocytogenes in Milk Samples by Signal Amplification Quartz Crystal Microbalance Sensor , 2010 .

[70]  M. Chiriacò,et al.  Impedance Sensing Platform for Detection of the Food Pathogen Listeria monocytogenes , 2018, Electronics.

[71]  X. D. Hoa,et al.  Microfluidic filtration and extraction of pathogens from food samples by hydrodynamic focusing and inertial lateral migration , 2015, Biomedical Microdevices.

[72]  M. Danyluk,et al.  Cross contamination of Escherichia coli O157:H7 between lettuce and wash water during home-scale washing. , 2015, Food microbiology.

[73]  Nicholas J. Ashbolt,et al.  Review of multi-criteria decision aid for integrated sustainability assessment of urban water systems , 2008 .

[74]  R. C. Whiting,et al.  A review of Listeria monocytogenes: An update on outbreaks, virulence, dose-response, ecology, and risk assessments , 2017 .

[75]  S. Hoffmann,et al.  Making Sense of Recent Cost-of-Foodborne-Illness Estimates , 2014 .

[76]  Yanbin Li,et al.  Interdigitated Array microelectrode-based electrochemical impedance immunosensor for detection of Escherichia coli O157:H7. , 2004, Analytical chemistry.

[77]  Dan Wang,et al.  Fast and sensitive detection of foodborne pathogen using electrochemical impedance analysis, urease catalysis and microfluidics. , 2016, Biosensors & bioelectronics.

[78]  Stefan Seeger,et al.  Understanding protein adsorption phenomena at solid surfaces. , 2011, Advances in colloid and interface science.