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

International Journal of Molecular Sciences

Ka L. Hong¹  Luisa Battistella¹  Alysia D. Salva¹  Ryan M. Williams¹  Letha J. Sooter¹ 

[1] Department of Pharmaceutical Sciences, West Virginia University, 1 Medical Center Drive, P.O. Box 9530, Morgantown, WV 26506, USA; E-Mails:

关键词: SELEX;    in vitro selection;    aptamer;    ssDNA;    molecular recognition element (MRE);    alpha toxin;    Staphylococcus aureus;    ELISA;   

DOI  :  10.3390/ijms16022794

来源: mdpi

【 摘 要 】

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 (K_d) 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.

【 授权许可】

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