Design and 3D printing of an electrochemical sensor for Listeria monocytogenes detection based on loop mediated isothermal amplification

Ane Rivas-Macho; Unai Eletxigerra; Ruth Diez-Ahedo; Santos Merino; Antton Sanjuan; M Mounir Bou-Ali; Leire Ruiz-Rubio; Javier Del Campo; José Luis Vilas-Vilela; Felipe Goñi-de-Cerio; Garbiñe Olabarria

doi:10.1016/j.heliyon.2022.e12637

Design and 3D printing of an electrochemical sensor for Listeria monocytogenes detection based on loop mediated isothermal amplification

Heliyon. 2022 Dec 26;9(1):e12637. doi: 10.1016/j.heliyon.2022.e12637. eCollection 2023 Jan.

Authors

Affiliations

¹ Gaiker, GAIKER Technology Centre, Basque Research and Technology Alliance, Zamudio 48170, Spain.
² Surface Chemistry and Nanotechnologies Unit, Tekniker, Eibar 20600, Spain.
³ Departamento de Electricidad y Electrónica, Universidad Del País Vasco UPV/EHU, 48940 Leioa, Spain.
⁴ Mechanical and Manufacturing Department, Mondragon University, Loramendi 4, Apdo. 23, 20500 Mondragon, Spain.
⁵ Macromolecular Chemistry Group, Department of Physical Chemistry, Faculty of Science and Technology, University of the Basque Country UPV/EHU, 48940 Leioa, Spain.
⁶ Basque Center for Materials, Applications and Nanostructures, Martina Casiano Building, UPV/EHU Science Park, 48940 Leioa, Spain.
⁷ IKERBASQUE, Basque Foundation for Science, Plaza Euskadi, 5, Bilbao 48009, Spain.

Abstract

The aim of this work is the design and 3D printing of a new electrochemical sensor for the detection of Listeria monocytogenes based on loop mediated isothermal amplification (LAMP). The food related diseases involve a serious health issue all over the world. Listeria monocytogenes is one of the major problems of contaminated food, this pathogen causes a disease called listeriosis with a high rate of hospitalization and mortality. Having a fast, sensitive and specific detection method for food quality control is a must in the food industry to avoid the presence of this pathogen in the food chain (raw materials, facilities and products). A point-of-care biosensor based in LAMP and electrochemical detection is one of the best options to detect the bacteria on site and in a very short period of time. With the numerical analysis of different geometries and flow rates during sample injection in order to avoid bubbles, an optimized design of the microfluidic biosensor chamber was selected for 3D-printing and experimental analysis. For the electrochemical detection, a novel custom gold concentric-3-electrode consisting in a working electrode, reference electrode and a counter electrode was designed and placed in the bottom of the chamber. The LAMP reaction was optimized specifically for a primers set with a limit of detection of 1.25 pg of genomic DNA per reaction and 100% specific for detecting all 12 Listeria monocytogenes serotypes and no other Listeria species or food-related bacteria. The methylene blue redox-active molecule was tested as the electrochemical transducer and shown to be compatible with the LAMP reaction and very clearly distinguished negative from positive food samples when the reaction is measured at the end-point inside the biosensor.

Keywords: 3D printing; Biosensor; Electrochemical detection; Listeria monocytogenes; Loop-mediated isothermal AMPlification (LAMP); Numerical analysis.