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Sensors 2015, 15(4), 8884-8897; doi:10.3390/s150408884

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
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Author to whom correspondence should be addressed.
Academic Editor: Stephane Evoy
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)
View Full-Text   |   Download PDF [1204 KB, uploaded 15 April 2015]   |  

Abstract

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
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. (CC BY 4.0).

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MDPI and ACS Style

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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