Abstract
The detection of mycotoxins is crucial because of their toxicity in plants, animals, and humans. It is very important to determine whether food products are contaminated with mycotoxins such as ochratoxin A (OTA), as mycotoxins can survive heat treatments and hydrolysis. In this study, we designed a fluorescence resonance energy transfer (FRET)-based system that exploits antibody-antigen binding to detect mycotoxins more rapidly and easily than other currently available methods. In addition, we were able to effectively counteract the matrix effect in the sample by using a nitrocellulose membrane that enabled fluorescence measurement in coffee samples. The developed FRET on lateral flow immunoassay (FRET-LFI) system was used to detect OTA at a limit of detection (LOD) of 0.64 ng∙mL−1, and the test can be completed in only 30 min. Moreover, OTA in coffee samples was successfully detected at a LOD of 0.88 ng∙mL−1, overcoming the matrix effect, owing to the chromatographic properties of the capillary force of the membrane. We believe that the developed system can be used as a powerful tool for the sensitive diagnosis of harmful substances such as mycotoxins and pesticides for environmental and food quality control monitoring.
Highlights
Mycotoxins are secondary metabolites produced by fungi that can cause diseases or abnormalities in humans and animals [1,2,3]
We aimed to achieve the following in our developed fluorescence resonance energy transfer (FRET)-lateral flow immunoassay (LFI) system: (1) a rapid and simple label-free immunoassay system; (2) a signal-on quantitative method for small molecule detection; and (3) a system in which the matrix effect is negated in order to apply this system directly to real samples, especially darkly colored solutions
The developed FRET-LFI in this study was designed based on the FRET between ochratoxin A (OTA) and the anti-OTA antibody
Summary
Mycotoxins are secondary metabolites produced by fungi that can cause diseases or abnormalities in humans and animals [1,2,3]. The toxic effect of these metabolites in mammals is known as mycotoxicosis [4,5]. Mycotoxins usually contaminate grains or nuts, which are useful hosts for the breeding of molds [6,7]; they are highly influenced by environmental factors such as temperature, moisture, and rainfall [8,9]. OTA has recently gained a lot of attention because of its biological toxicity—it can cause effects such as teratogenicity, carcinogenicity, immune toxicity, and even fertility inhibition [11,12]. OTA is related to a human disease known as Balkan endemic nephropathy, which is mainly found in Southeastern Europe [13]. The detection of OTA in food products is important because the ingestion of OTA-contaminated food constitutes a risk
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