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Open AccessArticle

Comparison of Whole-Cell SELEX Methods for the Identification of Staphylococcus Aureus-Specific DNA Aptamers

National Academy of Agricultural Science, 310 Nongsaengmyeng-ro, Wansan-gu, Jeonju 560500, Korea
Author to whom correspondence should be addressed.
Academic Editor: Stephane Evoy
Sensors 2015, 15(4), 8884-8897;
Received: 6 February 2015 / Revised: 28 March 2015 / Accepted: 9 April 2015 / Published: 15 April 2015
(This article belongs to the Special Issue Biosensors for Pathogen Detection)
Whole-cell Systemic Evolution of Ligands by Exponential enrichment (SELEX) is the process by which aptamers specific to target cells are developed. Aptamers selected by whole-cell SELEX have high affinity and specificity for bacterial surface molecules and live bacterial targets. To identify DNA aptamers specific to Staphylococcus aureus, we applied our rapid whole-cell SELEX method to a single-stranded ssDNA library. To improve the specificity and selectivity of the aptamers, we designed, selected, and developed two categories of aptamers that were selected by two kinds of whole-cell SELEX, by mixing and combining FACS analysis and a counter-SELEX process. Using this approach, we have developed a biosensor system that employs a high affinity aptamer for detection of target bacteria. FAM-labeled aptamer sequences with high binding to S. aureus, as determined by fluorescence spectroscopic analysis, were identified, and aptamer A14, selected by the basic whole-cell SELEX using a once-off FACS analysis, and which had a high binding affinity and specificity, was chosen. The binding assay was evaluated using FACS analysis. Our study demonstrated the development of a set of whole-cell SELEX derived aptamers specific to S. aureus; this approach can be used in the identification of other bacteria. View Full-Text
Keywords: aptamer; biosensor; pathogen detection; whole-cell SELEX aptamer; biosensor; pathogen detection; whole-cell SELEX
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Moon, J.; Kim, G.; Park, S.B.; Lim, J.; Mo, C. Comparison of Whole-Cell SELEX Methods for the Identification of Staphylococcus Aureus-Specific DNA Aptamers. Sensors 2015, 15, 8884-8897.

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