Aptamer-Based Assay For Detection Of Ochratoxin A

• Main Author: Bartley, Amanda Nicole
• Format: Others
• Published: FIU Digital Commons 2018
• Subjects: Ochratoxin A; mycotoxin; aptamer; fluorescence; assay; Analytical Chemistry; Biochemistry; Environmental Chemistry; Environmental Health; Toxicology
• Online Access: https://digitalcommons.fiu.edu/etd/3894; https://digitalcommons.fiu.edu/cgi/viewcontent.cgi?article=5141&context=etd

Ochratoxin A (OTA) is a potent mycotoxin found in a wide range of agricultural products that has been linked to mitochondrial damage and renal disease. The standard methods for OTA analysis currently rely on the use of high-performance liquid chromatography (HPLC) coupled to fluorescence detection or mass spectrometry. Toward a highthroughput analysis of OTA, a single-stranded DNA aptamer, modified with a fluorophore, coupled to a complementary sequence, modified with a FRET-based quencher that dissociates in the presence of the target toxin, is proposed. In order to integrate “target trapping,” aptamer immobilization methods were explored to mediate interference issues. Assays were evaluated using wine and blood serum matrices. A solution-based assay in a 96-well plate format provided a limit-of-detection of 2.7 ng/mL which would be suitable for many of the proposed applications. Immobilized aptamer formats, however, were not reliable, and a range of limitations to applications of the assay were identified.