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

Sensors

Jihea Moon²  Giyoung Kim¹  Saet Byeol Park²  Jongguk Lim²  Changyeun Mo² 

[1] National Academy of Agricultural Science, 310 Nongsaengmyeng-ro, Wansan-gu, Jeonju 560500, Korea;

关键词: aptamer;    biosensor;    pathogen detection;    whole-cell SELEX;   

DOI  :  10.3390/s150408884

来源: mdpi

【 摘 要 】

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.

【 授权许可】

CC BY   
© 2015 by the authors; licensee MDPI, Basel, Switzerland.