Citation: LIN Hao, HE Xuan, CHEN Yan, PANG Lu-Yi, NIU Lei, FU Xiu-Li. A Rapid Ratiometric Fluorescence Biosensor for Detection of Ochratoxin A Based on Paper Chip[J]. Chinese Journal of Analytical Chemistry, ;2022, 50(9): 1336-1344. doi: 10.19756/j.issn.0253-3820.221127 shu

A Rapid Ratiometric Fluorescence Biosensor for Detection of Ochratoxin A Based on Paper Chip

LIN Hao ¹ ,
HE Xuan ¹ ,
CHEN Yan ² ,
PANG Lu-Yi ¹ ,
NIU Lei ¹ ,
FU Xiu-Li ^1,,

1.
School of Chemistry and Chemical Engineering, Yantai University, Yantai 264005, China;
2.
School of Resources and Environmental Engineering, Shandong Agriculture and Engineering University, Jinan 250100, China

Corresponding author: FU Xiu-Li, fuxiuli@ytu.edu.cn
Received Date: 15 March 2022
Revised Date: 20 May 2022

Fund Project: Supported by the Qingchuang Science and Technology program of Shandong Province, China (No.2019KJF029) and the National Natural Science Foundation of China (Nos.61801274, 21705139).

A novel ratiometric fluorescent paper chip was constructed using dual fluorescent dyes based on fluorescence resonance energy transfer (FRET) between donor Cy3 and acceptor Cy5. The Cy3-labeled aptamer and Cy5-labeled auxiliary DNA (aDNA) were simultaneously combined with the complementary DNA (cDNA) pre-modified on the surface of paper chip to form a specific double stranded structure. After forming double stranded structure, a ratiometric fluorescence signal change was realized by FRET between donor Cy3 and acceptor Cy5. In this case, the fluorescence intensity of Cy3 decreased accompanied by the increasing of the fluorescence intensity of Cy5. In the presence of ochratoxin A (OTA), the specific binding of aptamer toward OTA led to Cy3-labeled aptamer detaching from the double stranded structure, which resulted in an increasing distance between Cy3 and Cy5. Therefore, the fluorescence intensity of Cy3 increased while the fluorescence intensity of Cy5 decreased. The fluorescence response ratio values (F₅₆₇/F₆₆₉:the fluorescence intensity of Cy3 at 567 nm to the fluorescence intensity of Cy5 at 669 nm) exhibited a good linear response with OTA concentration in the range of 10-300 nmol/L and the detection limit was 5.6 nmol/L. The designed ratiometric fluorescent paper chip was applied to detect OTA in spiked peanut and red wine samples, and the recoveries were 92.7%-107.6%. This sensor provided an efficient and convenient method for detection of mycotoxin in food.
- Paper chip,
- Ratiometric fluorescence,
- Fluorescence resonance energy transfer,
- Ochratoxin A,
- Aptamer
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