Citation: QU Yao, ZHANG Ya-Qi, XIAO Guang, YANG Cheng. Aptasensor Based on Nucleic Acid Base Quenching Fluorophore for Detection of Ochratoxin A[J]. Chinese Journal of Analytical Chemistry, ;2020, 48(10): 1409-1415. doi: 10.19756/j.issn.0253-3820.201260 shu

Aptasensor Based on Nucleic Acid Base Quenching Fluorophore for Detection of Ochratoxin A

State Key Laboratory of Fine Chemicals, School of Chemistry, College of Chemical Engineering, Dalian University of Technology, Dalian 116024, China

Received Date: 27 May 2020
Revised Date: 6 July 2020

Fund Project: This work was supported by the Fundamental Research Funds for the Central Universities (No. DUT15RC(3) 064),the Scientific Research Foundation for Doctoral Research of Liaoning Province, China (No. 201501167) and the Open Funds of the State Key Laboratory of Electroanalytical Chemistry (No. SKLEAC201604).

Ochratoxin A (OTA) is one kind of mycotoxin, which is toxic and widely distributed. To avoid the poison of OTA, it is necessary to develop a rapid, low-cost and highly sensitive detection method to evaluate the concentration of OTA in the food. Here, an aptamer biosensor was developed to detect OTA in wine using single labeled oligonucleotide with fluorescein (FAM) as probe and guanine in the aptamer as quencher. Guanine was a heterocyclic compound that had a high electron density and the lowest oxidation potential in nucleic acid bases. Guanine was easily to be oxidized. As an electron donor, the fluorophore could be quenched by guanine through the process of photoinduced electron transfer (PET). When the probe labeled with FAM hybridized with the OTA aptamer, the fluorescence of FAM was quenched by the guanine in OTA aptamer. In the presence of OTA, due to the specific interaction between OTA and aptamer, OTA induced aptamer folding to G-quadruplexes, which inhibited the binding of probe with aptamer, and thus led to the fluorescence intensity recovery. The concentration of OTA could be determined by the recovery ratio of FAM fluorescence intensity. The linear range for detection of OTA by this aptasenor was 0.67-7.80 nmol/L, the linear calibration equation was R(%)=-0.02+1.76C_OTA (nmol/L), and LOD was 0.67 nmol/L (0.27 g/kg, S/N=3).The recoveries of OTA in red wine sample were in the range of 92.4%-100.9%. Compared with sensing strategies based on gold nanoparticles, single-walled nanotubes and graphene oxide, this method had many advantages such as low cost, good selectivity and high sensitivity.
- Guanine base,
- Fluorescence quenching,
- Aptamer,
- Single-label fluorescein,
- Ochratoxin A
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