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Int. J. Mol. Sci. 2015, 16(2), 2794-2809; doi:10.3390/ijms16022794

In Vitro Selection of Single-Stranded DNA Molecular Recognition Elements against S. aureus Alpha Toxin and Sensitive Detection in Human Serum

1
Department of Pharmaceutical Sciences, West Virginia University, 1 Medical Center Drive, P.O. Box 9530, Morgantown, WV 26506, USA
2
Memorial Sloan Kettering Cancer Center, Molecular Pharmacology and Chemistry, 1275 York Avenue, P.O. Box 425, New York, NY 10065, USA
*
Author to whom correspondence should be addressed.
Academic Editor: Ritva Tikkanen
Received: 18 November 2014 / Revised: 5 December 2014 / Accepted: 19 January 2015 / Published: 27 January 2015
(This article belongs to the Section Molecular Recognition)
View Full-Text   |   Download PDF [1666 KB, uploaded 27 January 2015]   |  

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. View Full-Text
Keywords: SELEX; in vitro selection; aptamer; ssDNA; molecular recognition element (MRE); alpha toxin; Staphylococcus aureus; ELISA SELEX; in vitro selection; aptamer; ssDNA; molecular recognition element (MRE); alpha toxin; Staphylococcus aureus; ELISA
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

Hong, K.L.; Battistella, L.; Salva, A.D.; Williams, R.M.; Sooter, L.J. In Vitro Selection of Single-Stranded DNA Molecular Recognition Elements against S. aureus Alpha Toxin and Sensitive Detection in Human Serum. Int. J. Mol. Sci. 2015, 16, 2794-2809.

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Int. J. Mol. Sci. EISSN 1422-0067 Published by MDPI AG, Basel, Switzerland RSS E-Mail Table of Contents Alert
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