A Novel Fluorescent Aptasensor Based on Dual-labeled DNA Nanostructure for Simultaneous Detection of Ochratoxin A and Aflatoxin B1

Zhao, Luyang; Liang, Xiujun; Liu, Yong; Wei, Min; Jin, Huali

doi:10.1007/s10895-022-03071-5

Cited by 3 publications

(2 citation statements)

References 48 publications

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“…In this assay, a specific FAM-labeled truncated 26-mer aptamer (5′-FAM-ATCACGTGTT-GTCTCTCTGTGTCTCGTG-3′) with a stem-loop secondary structure stabilized by four base pairs was used. To detect AFB1, ssDNA complementary to either the 5′ or 3′ end of the aptamer was typically used to ensure the proximity between the quencher and the fluorophore [32][33][34][35][36]. Considering the flexibility of ssDNAs [37], we applied an ssDNA complementary to the loop region of the aptamer (Figure 1b) identified in previous studies as the binding site for AFB1 [28].…”

Section: Principle Of Afb1 Detectionmentioning

confidence: 99%

“…In addition, donor-acceptor pairs based on organic compounds, namely fluorescein-BHQ-1 [31], fluorescein-DABCYL [32], as well as binary systems of Cy5-BHQ-2 and Cy3-Cy5 fluorophores [33], have been implemented. More complex systems using a pair of FAM/BHQ-1 labeled ssDNA with non-overlapping complementary sites on the aptamer [34] or a Cy5-BHQ-2/Cy3-BHQ-2 pair [35] have also been characterized.…”

Section: Introductionmentioning

confidence: 99%

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A Fluorescence Resonance Energy Transfer Aptasensor for Aflatoxin B1 Based on Ligand-Induced ssDNA Displacement

Serebrennikova,

Samokhvalov,

Zherdev

et al. 2023

Molecules

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In this study, a fluorescence resonance energy transfer (FRET)-based aptasensor for the detection of aflatoxin B1 (AFB1) was designed using a carboxyfluorescein (FAM)-labeled aptamer and short complementary DNA (cDNA) labeled with low molecular quencher RTQ1. The sensing principle was based on the detection of restored FAM-aptamer fluorescence due to the ligand-induced displacement of cDNA in the presence of AFB1, leading to the destruction of the aptamer/cDNA duplex and preventing the convergence of FAM and RTQ1 at the effective FRET distance. Under optimal sensing conditions, a linear correlation was obtained between the fluorescence intensity of the FAM-aptamer and the AFB1 concentration in the range of 2.5–208.3 ng/mL with the detection limit of the assay equal to 0.2 ng/mL. The assay time was 30 min. The proposed FRET aptasensor has been successfully validated by analyzing white wine and corn flour samples, with recovery ranging from 76.7% to 91.9% and 84.0% to 86.5%, respectively. This work demonstrates the possibilities of labeled cDNA as an effective and easily accessible tool for sensitive AFB1 detection. The homogeneous FRET aptasensor is an appropriate choice for contaminant screening in complex matrices.

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