title: In vitro selection of single-stranded DNA molecular recognition elements against S. aureus alpha toxin and sensitive detection in human serum.
abstract: Alpha toxin is one of the major virulence factors secreted by Staphylococcus aureus, a bacterium that is responsible for a wide variety of infections in both community and hospital settings. Due to the prevalence of S. aureus related infections and the emergence of methicillin-resistant S. aureus, rapid and accurate diagnosis of S. aureus infections is crucial in benefiting patient health outcomes. In this study, a rigorous Systematic Evolution of Ligands by Exponential Enrichment (SELEX) variant previously developed by our laboratory was utilized to select a single-stranded DNA molecular recognition element (MRE) targeting alpha toxin with high affinity and specificity. At the end of the 12-round selection, the selected MRE had an equilibrium dissociation constant (Kd) of 93.7 ± 7.0 nM. Additionally, a modified sandwich enzyme-linked immunosorbent assay (ELISA) was developed by using the selected ssDNA MRE as the toxin-capturing element and a sensitive detection of 200 nM alpha toxin in undiluted human serum samples was achieved.
authors: Department of Pharmaceutical Sciences, West Virginia University, 1 Medical Center Drive, P.O. Box 9530, Morgantown, WV 26506, USA. khong@mix.wvu.edu.
authors: Ka L
authors: KL
authors: Hong
authors: Department of Pharmaceutical Sciences, West Virginia University, 1 Medical Center Drive, P.O. Box 9530, Morgantown, WV 26506, USA. lbattistella@mail.wvu.edu.
authors: Luisa
authors: L
authors: Battistella
authors: Department of Pharmaceutical Sciences, West Virginia University, 1 Medical Center Drive, P.O. Box 9530, Morgantown, WV 26506, USA. asalva@mix.wvu.edu.
authors: Alysia D
authors: AD
authors: Salva
authors: Department of Pharmaceutical Sciences, West Virginia University, 1 Medical Center Drive, P.O. Box 9530, Morgantown, WV 26506, USA. rwilliams@mix.wvu.edu.
authors: Ryan M
authors: RM
authors: Williams
authors: Department of Pharmaceutical Sciences, West Virginia University, 1 Medical Center Drive, P.O. Box 9530, Morgantown, WV 26506, USA. lsooter@hsc.wvu.edu.
authors: Letha J
authors: LJ
authors: Sooter
mesh_terms: D001427:Bacterial Toxins / Q000097:blood*; D001483:Base Sequence; D004277:DNA, Single-Stranded / Q000737:chemistry / Q000378:metabolism*; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D007700:Kinetics; D009690:Nucleic Acid Conformation; D052156:SELEX Aptamer Technique; D013211:Staphylococcus aureus / Q000378:metabolism*; D020349:Surface Plasmon Resonance
chemical_list: D001427:Bacterial Toxins; D004277:DNA, Single-Stranded
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