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Fast and Accurate Determination of Minute Ochratoxin A Levels in Cereal Flours and Wine with the Label-Free White Light Reflectance Spectroscopy Biosensing Platform †

Ochratoxin A (OTA) is one of the most toxic naturally encountered contaminants and is found in a variety of foods and beverages, including cereals and wine. Driven by the strict regulations regarding the maximum allowable OTA concentration in foodstuff and the necessity for on-site determination, th...

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Autores principales: Karachaliou, Chrysoula-Evangelia, Koukouvinos, Georgios, Zisis, Grigoris, Kizis, Dimosthenis, Krystalli, Evangelia, Siragakis, George, Goustouridis, Dimitris, Kakabakos, Sotirios, Petrou, Panagiota, Livaniou, Evangelia, Raptis, Ioannis
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9599867/
https://www.ncbi.nlm.nih.gov/pubmed/36291014
http://dx.doi.org/10.3390/bios12100877
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author Karachaliou, Chrysoula-Evangelia
Koukouvinos, Georgios
Zisis, Grigoris
Kizis, Dimosthenis
Krystalli, Evangelia
Siragakis, George
Goustouridis, Dimitris
Kakabakos, Sotirios
Petrou, Panagiota
Livaniou, Evangelia
Raptis, Ioannis
author_facet Karachaliou, Chrysoula-Evangelia
Koukouvinos, Georgios
Zisis, Grigoris
Kizis, Dimosthenis
Krystalli, Evangelia
Siragakis, George
Goustouridis, Dimitris
Kakabakos, Sotirios
Petrou, Panagiota
Livaniou, Evangelia
Raptis, Ioannis
author_sort Karachaliou, Chrysoula-Evangelia
collection PubMed
description Ochratoxin A (OTA) is one of the most toxic naturally encountered contaminants and is found in a variety of foods and beverages, including cereals and wine. Driven by the strict regulations regarding the maximum allowable OTA concentration in foodstuff and the necessity for on-site determination, the development of fast and sensitive methods for the OTA determination in cereal flours and wine samples, based on white light reflectance spectroscopy, is presented. The method relied on appropriately engineered silicon chips, on top of which an OTA-protein conjugate was immobilized. A polyclonal antibody against OTA was then employed to detect the analyte in the framework of a competitive immunoassay; followed by the subsequent addition of a biotinylated secondary antibody and streptavidin for signal enhancement. A small size instrument performed all assay steps automatically and the bioreactions were monitored in real time as the software converted the spectral shifts into effective biomolecular adlayer thickness increase. The assay developed had a detection limit of 0.03 ng/mL and a working range up to 200 ng/mL. The assay lasted 25 min (less than 1h, including calibrators/antibody pre-incubation) and was accomplished following a simple sample preparation protocol. The method was applied to corn and wheat flour samples and white and red wines with recovery values ranging from 87.2 to 111%. The simplicity of the overall assay protocol and convenient instrumentation demonstrates the potential of the immunosensor developed for OTA detection at the point of need.
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spelling pubmed-95998672022-10-27 Fast and Accurate Determination of Minute Ochratoxin A Levels in Cereal Flours and Wine with the Label-Free White Light Reflectance Spectroscopy Biosensing Platform † Karachaliou, Chrysoula-Evangelia Koukouvinos, Georgios Zisis, Grigoris Kizis, Dimosthenis Krystalli, Evangelia Siragakis, George Goustouridis, Dimitris Kakabakos, Sotirios Petrou, Panagiota Livaniou, Evangelia Raptis, Ioannis Biosensors (Basel) Article Ochratoxin A (OTA) is one of the most toxic naturally encountered contaminants and is found in a variety of foods and beverages, including cereals and wine. Driven by the strict regulations regarding the maximum allowable OTA concentration in foodstuff and the necessity for on-site determination, the development of fast and sensitive methods for the OTA determination in cereal flours and wine samples, based on white light reflectance spectroscopy, is presented. The method relied on appropriately engineered silicon chips, on top of which an OTA-protein conjugate was immobilized. A polyclonal antibody against OTA was then employed to detect the analyte in the framework of a competitive immunoassay; followed by the subsequent addition of a biotinylated secondary antibody and streptavidin for signal enhancement. A small size instrument performed all assay steps automatically and the bioreactions were monitored in real time as the software converted the spectral shifts into effective biomolecular adlayer thickness increase. The assay developed had a detection limit of 0.03 ng/mL and a working range up to 200 ng/mL. The assay lasted 25 min (less than 1h, including calibrators/antibody pre-incubation) and was accomplished following a simple sample preparation protocol. The method was applied to corn and wheat flour samples and white and red wines with recovery values ranging from 87.2 to 111%. The simplicity of the overall assay protocol and convenient instrumentation demonstrates the potential of the immunosensor developed for OTA detection at the point of need. MDPI 2022-10-15 /pmc/articles/PMC9599867/ /pubmed/36291014 http://dx.doi.org/10.3390/bios12100877 Text en © 2022 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Karachaliou, Chrysoula-Evangelia
Koukouvinos, Georgios
Zisis, Grigoris
Kizis, Dimosthenis
Krystalli, Evangelia
Siragakis, George
Goustouridis, Dimitris
Kakabakos, Sotirios
Petrou, Panagiota
Livaniou, Evangelia
Raptis, Ioannis
Fast and Accurate Determination of Minute Ochratoxin A Levels in Cereal Flours and Wine with the Label-Free White Light Reflectance Spectroscopy Biosensing Platform †
title Fast and Accurate Determination of Minute Ochratoxin A Levels in Cereal Flours and Wine with the Label-Free White Light Reflectance Spectroscopy Biosensing Platform †
title_full Fast and Accurate Determination of Minute Ochratoxin A Levels in Cereal Flours and Wine with the Label-Free White Light Reflectance Spectroscopy Biosensing Platform †
title_fullStr Fast and Accurate Determination of Minute Ochratoxin A Levels in Cereal Flours and Wine with the Label-Free White Light Reflectance Spectroscopy Biosensing Platform †
title_full_unstemmed Fast and Accurate Determination of Minute Ochratoxin A Levels in Cereal Flours and Wine with the Label-Free White Light Reflectance Spectroscopy Biosensing Platform †
title_short Fast and Accurate Determination of Minute Ochratoxin A Levels in Cereal Flours and Wine with the Label-Free White Light Reflectance Spectroscopy Biosensing Platform †
title_sort fast and accurate determination of minute ochratoxin a levels in cereal flours and wine with the label-free white light reflectance spectroscopy biosensing platform †
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9599867/
https://www.ncbi.nlm.nih.gov/pubmed/36291014
http://dx.doi.org/10.3390/bios12100877
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