Fluorometric aptamer based assay for ochratoxin A based on the use of exonuclease III
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This study describes an aptamer based assay for the mycotoxin ochratoxin A (OTA). The method is based on the use of an OTA-specific aptamer, exonuclease (Exo) III, SYBR Gold as a fluorescent probe, and a complementary strand that specifically combines with the aptamer. In the presence of OTA, the aptamer and OTA hybridize, thereby resulting in the formation of ssDNA, which is not digested by Exo III. Intense fluorescence is observed after addition of SYBR Gold (best measured at excitation/emission wavelengths of 495/540 nm). Fluorescence increases linearly with the log of the OTA concentration in the range from 8 to 1000 ng·mL−1. The detection limit is 4.7 ng·mL−1. The assay was applied to the determination of OTA in diluted [2%(v/v)] red wine, and recoveries and RSDs ranged between 93.5% and 113.8%, and between 3.2% and 5.7%, respectively.
KeywordsMycotoxin Fluorescent probe Exo III Complementary strand
This work was supported by the National Natural Science Foundation of China (Nos. 31460423 and 31360384), the department of Sciences & Technology of Jilin Province (20160520047JH) and the department of education of Jilin Province (2016252).
Compliance with ethical standards
The author(s) declare that they have no competing interests.
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