Aptamer-Based Assay For Detection Of Ochratoxin A

Authors

Amanda Nicole BartleyFollow

Document Type

Thesis

Degree

Master of Science (MS)

Major/Program

Forensic Science

First Advisor's Name

John Berry

First Advisor's Committee Title

Committee chair

Second Advisor's Name

DeEtta Mills

Second Advisor's Committee Title

Committee member

Third Advisor's Name

Yi Xiao

Third Advisor's Committee Title

Committee member

Keywords

Ochratoxin A, mycotoxin, aptamer, fluorescence, assay

Date of Defense

11-8-2018

Abstract

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.

Identifier

FIDC007013

ORCID

https://orcid.org/0000-0001-6877-0165

Recommended Citation

Bartley, Amanda Nicole, "Aptamer-Based Assay For Detection Of Ochratoxin A" (2018). FIU Electronic Theses and Dissertations. 3894.
https://digitalcommons.fiu.edu/etd/3894