Selection and Characterization of a Novel DNA Aptamer for Label-Free Fluorescence Biosensing of Ochratoxin A
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...
| Main Authors: | , , , , , |
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| Format: | Article |
| Language: | English |
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MDPI AG
2014-08-01
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| Series: | Toxins |
| Subjects: | |
| Online Access: | http://www.mdpi.com/2072-6651/6/8/2435 |
| _version_ | 1798034877504815104 |
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| author | Maureen McKeague Ranganathan Velu Kayla Hill Viola Bardóczy Tamás Mészáros Maria C. DeRosa |
| author_facet | Maureen McKeague Ranganathan Velu Kayla Hill Viola Bardóczy Tamás Mészáros Maria C. DeRosa |
| author_sort | Maureen McKeague |
| collection | DOAJ |
| description | 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. |
| first_indexed | 2024-04-11T20:49:22Z |
| format | Article |
| id | doaj.art-b80ba8bcb752495c85572bba78d00e6b |
| institution | Directory Open Access Journal |
| issn | 2072-6651 |
| language | English |
| last_indexed | 2024-04-11T20:49:22Z |
| publishDate | 2014-08-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Toxins |
| spelling | doaj.art-b80ba8bcb752495c85572bba78d00e6b2022-12-22T04:03:53ZengMDPI AGToxins2072-66512014-08-01682435245210.3390/toxins6082435toxins6082435Selection and Characterization of a Novel DNA Aptamer for Label-Free Fluorescence Biosensing of Ochratoxin AMaureen McKeague0Ranganathan Velu1Kayla Hill2Viola Bardóczy3Tamás Mészáros4Maria C. DeRosa5Department of Bioengineering, Stanford University, 443 Via Ortega, MC 4245, Stanford, CA 94305, USAChemistry Department, Carleton University, 1125 Colonel By Drive, Ottawa, ON K1S 5B6, CanadaChemistry Department, Carleton University, 1125 Colonel By Drive, Ottawa, ON K1S 5B6, CanadaDepartment of Applied Biotechnology and Food Science, Budapest University of Technology Economics, Szt. Gellért tér 4, H-1111 Budapest, HungaryDepartment of Medical Chemistry, Molecular Biology and Pathobiochemistry, Semmelweis University, Tûzoltó u. 37-47, H-1094 Budapest, HungaryChemistry Department, Carleton University, 1125 Colonel By Drive, Ottawa, ON K1S 5B6, CanadaNucleic 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.http://www.mdpi.com/2072-6651/6/8/2435aptamerochratoxin Amycotoxinsin vitro selectionSELEXSYBR Green Ifluorescent assaybiosensingaptasensor |
| spellingShingle | Maureen McKeague Ranganathan Velu Kayla Hill Viola Bardóczy Tamás Mészáros Maria C. DeRosa Selection and Characterization of a Novel DNA Aptamer for Label-Free Fluorescence Biosensing of Ochratoxin A Toxins aptamer ochratoxin A mycotoxins in vitro selection SELEX SYBR Green I fluorescent assay biosensing aptasensor |
| title | Selection and Characterization of a Novel DNA Aptamer for Label-Free Fluorescence Biosensing of Ochratoxin A |
| title_full | Selection and Characterization of a Novel DNA Aptamer for Label-Free Fluorescence Biosensing of Ochratoxin A |
| title_fullStr | Selection and Characterization of a Novel DNA Aptamer for Label-Free Fluorescence Biosensing of Ochratoxin A |
| title_full_unstemmed | Selection and Characterization of a Novel DNA Aptamer for Label-Free Fluorescence Biosensing of Ochratoxin A |
| title_short | Selection and Characterization of a Novel DNA Aptamer for Label-Free Fluorescence Biosensing of Ochratoxin A |
| title_sort | selection and characterization of a novel dna aptamer for label free fluorescence biosensing of ochratoxin a |
| topic | aptamer ochratoxin A mycotoxins in vitro selection SELEX SYBR Green I fluorescent assay biosensing aptasensor |
| url | http://www.mdpi.com/2072-6651/6/8/2435 |
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