Specific and Simultaneous Detection of E. coli O157:H7 and Shiga-like Toxins Using a Label-Free Photonic Immunosensor

Author:

Fernández Ana1,Hernández Manuel2ORCID,Moreno Yolanda3ORCID,García-Hernández Jorge2ORCID

Affiliation:

1. Lumensia Sensors S.L., 46020 Valencia, Spain

2. Centro Avanzado de Microbiología de Alimentos, Biotechnology Department, Universitat Politècnica de València, 46022 Valencia, Spain

3. Instituto de Ingeniería de Agua y del Medioambiente, Universitat Politècnica de València, 46022 Valencia, Spain

Abstract

The current study outlines the advancement of an innovative technique for the simultaneous detection of E. coli O157:H7 and its Shiga-like toxins in food samples by utilizing a photonic label-free biosensor coupled with a microfluidic system. This detection method relies on ring resonator transduction that is functionalized with specific bioreceptors against O157:H7 on silicon nitride surfaces capable of binding specifically to the antigen bacterium and its verotoxins. This experiment included the characterization of selected monoclonal and polyclonal antibodies employed as detection probes through ELISA immunoassays exposed to target bacterial antigens. A thorough validation of photonic immunosensor detection was conducted on inoculated minced beef samples using reference standards for E. coli O157:H7 and its verotoxins (VTx1 and VTx2) and compared to gold-standard quantification. The lowest limit-of-detection values of 10 CFU/mL and 1 ppm were achieved for the detection of bacteria and its verotoxins. In this study, the lowest limit of quantification (LoQ) achieved for bacterial quantification was 100 CFU/mL, and, for verotoxins, it was 2 ppm. This study confirmed the effectiveness of a new quality control and food hygiene method, demonstrating the rapid and sensitive detection of E. coli O157:H7 and its verotoxins. This innovative approach has the potential to be applied in food production environments.

Funder

AVI

European Union

Publisher

MDPI AG

Reference76 articles.

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