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Anal Chim Acta ; 1272: 341503, 2023 Sep 01.
Artigo em Inglês | MEDLINE | ID: mdl-37355335

RESUMO

Ochratoxin A (OTA), a typical mycotoxin contaminant found in various agricultural products and foods, poses a serious threat to human health. In this study, an aptasensor based on a novel fluorescence probe comprising a G-rich DNA sequence (G43) and thioflavin T (ThT) was designed via hybridization chain reaction (HCR) for the ultrasensitive detection of OTA. G43 is a concatemer of G-quadruplex and G-triplex (a G-quadruplex-like structure with one G-quartet removed), which can drastically enhance the fluorescence intensity of ThT. For this strategy to work, the OTA aptamer is pro-locked in a hairpin structure, denoted "hairpin-locked aptamer" (HL-Apt). OTA binds to HL-Apt, opens the hairpin structure, releases the trigger sequence, and initiates the HCR reaction to form a long DNA duplex and numerous side chains. The side chains combine entirely with the complementary DNA and liberate the pro-locked G43 DNA, dramatically enhancing the intensity of the ThT fluorescence signal. The fluorescence intensity correlates linearly with the OTA concentration between 0.02 and 2.00 ng mL-1, and the method has a detection limit of 0.008 ng mL-1. The developed aptasensor was used to detect OTA in foodstuffs with satisfactory results.


Assuntos
Aptâmeros de Nucleotídeos , Técnicas Biossensoriais , Ocratoxinas , Humanos , Corantes Fluorescentes/química , Ocratoxinas/análise , Hibridização de Ácido Nucleico , DNA/genética , DNA/química , Aptâmeros de Nucleotídeos/química , Técnicas Biossensoriais/métodos , Limite de Detecção
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