title: Comparison of whole-cell SELEX methods for the identification of Staphylococcus aureus-specific DNA aptamers.
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.
authors.affiliation: National Academy of Agricultural Science, 310 Nongsaengmyeng-ro, Wansan-gu, Jeonju 560500, Korea. mmir95@gmail.com.
authors.forename: Jihea
authors.initials: J
authors.lastname: Moon
authors.affiliation: National Academy of Agricultural Science, 310 Nongsaengmyeng-ro, Wansan-gu, Jeonju 560500, Korea. giyoung@korea.kr.
authors.forename: Giyoung
authors.initials: G
authors.lastname: Kim
authors.affiliation: National Academy of Agricultural Science, 310 Nongsaengmyeng-ro, Wansan-gu, Jeonju 560500, Korea. veryvenus0911@gmail.com.
authors.forename: Saet Byeol
authors.initials: SB
authors.lastname: Park
authors.affiliation: National Academy of Agricultural Science, 310 Nongsaengmyeng-ro, Wansan-gu, Jeonju 560500, Korea. limjg@korea.kr.
authors.forename: Jongguk
authors.initials: J
authors.lastname: Lim
authors.affiliation: National Academy of Agricultural Science, 310 Nongsaengmyeng-ro, Wansan-gu, Jeonju 560500, Korea. cymoh100@korea.kr.
authors.forename: Changyeun
authors.initials: C
authors.lastname: Mo
mesh_terms: D052157:Aptamers, Nucleotide / Q000235:genetics*; D015374:Biosensing Techniques / Q000379:methods*; D052156:SELEX Aptamer Technique / Q000379:methods*; D013211:Staphylococcus aureus / Q000235:genetics*
chemical_list: D052157:Aptamers, Nucleotide