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Toxins 2014, 6(8), 2435-2452; doi:10.3390/toxins6082435
Article

Selection and Characterization of a Novel DNA Aptamer for Label-Free Fluorescence Biosensing of Ochratoxin A

1,2
, 2
, 2
, 3
, 4,5,*  and 2,*
1 Department of Bioengineering, Stanford University, 443 Via Ortega, MC 4245, Stanford, CA 94305, USA 2 Chemistry Department, Carleton University, 1125 Colonel By Drive, Ottawa, ON K1S 5B6, Canada 3 Department of Applied Biotechnology and Food Science, Budapest University of Technology Economics, Szt. Gellért tér 4, H-1111 Budapest, Hungary 4 Department of Medical Chemistry, Molecular Biology and Pathobiochemistry, Semmelweis University, Tûzoltó u. 37-47, H-1094 Budapest, Hungary 5 MTA-BME Technical Analytical Chemistry Research Group of the Hungarian Academy of Sciences, Szent Gellért tér 4, H-1111 Budapest, Hungary
* Authors to whom correspondence should be addressed.
Received: 16 June 2014 / Revised: 25 July 2014 / Accepted: 30 July 2014 / Published: 15 August 2014
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Abstract

Nucleic acid aptamers are emerging as useful molecular recognition tools for food safety monitoring. However, practical and technical challenges limit the number and diversity of available aptamer probes that can be incorporated into novel sensing schemes. This work describes the selection of novel DNA aptamers that bind to the important food contaminant ochratoxin A (OTA). Following 15 rounds of in vitro selection, sequences were analyzed for OTA binding. Two of the isolated aptamers demonstrated high affinity binding and selectivity to this mycotoxin compared to similar food adulterants. These sequences, as well as a truncated aptamer (minimal sequence required for binding), were incorporated into a SYBR® Green I fluorescence-based OTA biosensing scheme. This label-free detection platform is capable of rapid, selective, and sensitive OTA quantification with a limit of detection of 9 nM and linear quantification up to 100 nM.
Keywords: aptamer; ochratoxin A; mycotoxins; in vitro selection; SELEX; SYBR Green I; fluorescent assay; biosensing; aptasensor aptamer; ochratoxin A; mycotoxins; in vitro selection; SELEX; SYBR Green I; fluorescent assay; biosensing; aptasensor
This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

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McKeague, M.; Velu, R.; Hill, K.; Bardóczy, V.; Mészáros, T.; DeRosa, M.C. Selection and Characterization of a Novel DNA Aptamer for Label-Free Fluorescence Biosensing of Ochratoxin A. Toxins 2014, 6, 2435-2452.

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