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TitleAptamers For Detection of Staphylococcal Enterotoxins in Food Samples
Author(s)Oliveira, Ricardo
Pinho, Eva
DeStefano, Jeffrey
Azevedo, Nuno F.
Almeida, Carina
Issue date18-Nov-2021
CitationOliveira, Ricardo; Pinho, Eva; DeStefano, Jeffrey; Azevedo, Nuno F.; Almeida, Carina, Aptamers For Detection of Staphylococcal Enterotoxins in Food Samples. Dare2Change - Innovation-Driven Agrifood Business. Porto, Portugal, Nov 18-19, 62, 2021.
Abstract(s)Staphylococcal food poisoning is a gastrointestinal disease caused by the consumption of food containing toxins pre-formed by enterotoxigenic Staphylococcus. Current detection of these toxins relies on time-consuming antibody-based immunoassays associated with cross-reactivity, low sensitivity, and interference from food matrices [1]. Aptamers are single-stranded oligonucleotides with a defined three- dimensional shape that bind with high-affinity to a target molecule. They are selected by the Systematic Evolution of Ligands by Exponential Enrichment (SELEX) method which consists in the screening of a random oligonucleotide library down to highly specific sequences for the target, by the repetition of successive steps of selection, amplification, and conditioning. Applying unnatural nucleotides is an essential evolution of aptamers, considering the limitations in the chemical and biological stability of traditional DNA and RNA aptamers [2]. In this work, a SELEX procedure was applied to isolate aptamers with unnatural nucleotides (2-deoxy- 2-fluoroarabinonucleotides (FANA)) specific to staphylococcal enterotoxin A (SEA). After performing 12 rounds of selection, 24 potential FANA aptamers for SEA were identified. Although the nucleotide comparison shows no consensus sequences between selected oligonucleotides, the analysis of secondary structures reveals enrichment in similarly folding patterns. These results reinforce the potential of applying unnatural nucleotides in SELEX procedures and suggest that, for SEA, aptamer selection points to preferential secondary structures which are like other DNA aptamers already described [3]. This work was the start point to create a platform for the development of nucleic acid mimics aptamers that combines aptamers biorecognition ability and unnatural nucleotides diversity/increased affinity. The selected aptamers will then be incorporated into a portable assay that can be easily implemented at the food industry; and tested in real food matrices for validation and comparison with standard detection systems. Although it has been developed for SEA, this technological platform might be easily adapted to any food poisoning toxin or protein.
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AccessOpen access
Appears in Collections:CEB - Resumos em Livros de Atas / Abstracts in Proceedings

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