Newly Developed System for the Robust Detection of Listeria monocytogenes Based on a Bioelectric Cell Biosensor.

Agni Hadjilouka,Antonios Inglezakis,Theofylaktos Apostolou,Lazaros Dougiakis,Dimitrios Tsaltas,Konstantinos Loizou

doi:10.3390/bios10110178

Agni Hadjilouka, Antonios Inglezakis + Show 4 more

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https://doi.org/10.3390/bios10110178

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Abstract

Human food-borne diseases caused by pathogenic bacteria have been significantly increased in the last few decades causing numerous deaths worldwide. The standard analyses used for their detection have significant limitations regarding cost, special facilities and equipment, highly trained staff, and a long procedural time that can be crucial for foodborne pathogens with high hospitalization and mortality rates, such as Listeria monocytogenes. This study aimed to develop a biosensor that could detect L. monocytogenes rapidly and robustly. For this purpose, a cell-based biosensor technology based on the Bioelectric Recognition Assay (BERA) and a portable device developed by EMBIO Diagnostics, called B.EL.D (Bio Electric Diagnostics), were used. Membrane engineering was performed by electroinsertion of Listeria monocytogenes homologous antibodies into the membrane of African green monkey kidney (Vero) cells. The newly developed biosensor was able to detect the pathogen’s presence rapidly (3 min) at concentrations as low as 102 CFU mL−1, demonstrating a higher sensitivity than most existing biosensor-based methods. In addition, lack of cross-reactivity with other Listeria species, as well as with Escherichia coli, was shown, thus, indicating biosensor’s significant specificity against L. monocytogenes.

Highlights

Foodborne diseases are of great concern worldwide, as they cause thousands of deaths each year, as well as significant malfunctions in health care systems, national economies, and global trade.Bacteria, viruses, parasites, and chemicals that contaminate food at any stage of food production are the causative agents for these infections
We report the development of a portable Bioelectric Recognition Assay (BERA)-type sensor based on mammalian cells for the rapid detection of L. monocytogenes
Monoclonal antibodies against L. monocytogenes were purchased from antibodies-online.com and L. monocytogenes NCTC 11,994 and L. innocua NCTC 11,288 from Merck (Darmstadt, Germany)

Summary

Introduction

Viruses, parasites, and chemicals that contaminate food at any stage of food production are the causative agents for these infections. It is estimated that 600 million people (i.e., 10% of the global population) get sick and 420,000 people die every year due to the consumption of contaminated food [1]. Listeria monocytogenes is an intracellular pathogenic bacterium widely distributed in the environment that has been determined as a causative agent of serious epidemic and sporadic food-borne illnesses in humans. Listeriosis can lead to gastroenteritis, meningitis, or other severe symptoms with high hospitalization and mortality rates (20–30%), especially in vulnerable populations. In 2016, listeriosis was the most severe illness with the highest hospitalization and mortality rate in Europe

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