Selection of Single-Stranded DNA Molecular Recognition Elements against Exotoxin A Using a Novel Decoy-SELEX Method and Sensitive Detection of Exotoxin A in Human Serum.

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  • Additional Information
    • Affiliation:
      Department of Pharmaceutical Sciences, West Virginia University, 1 Medical Center Drive, P.O. Box 9530, Morgantown, WV 20506, USA
      School of Medicine, WV University, Morgantown, WV 20506, USA
      Department of Medical Laboratory Science, West Virginia University, Morgantown, WV 20506, USA
      Molecular Pharmacology & Chemistry Program, Memorial Sloan Kettering Cancer Center, 1275 York Avenue, New York, NY 10065, USA
      College of Pharmacy, Nova Southeastern University, Ft. Lauderdale, FL 33328, USA
    • Subject Terms:
    • Subject Terms:
    • Abstract:
      Exotoxin A is one of the virulence factors of Pseudomonas aeruginosa, a bacterium that can cause infections resulting in adverse health outcomes and increased burden to health care systems. Current methods of diagnosing P. aeruginosa infections are time consuming and can require significant preparation of patient samples. This study utilized a novel variation of the Systematic Evolution of Ligand by Exponential Enrichment, Decoy-SELEX, to identify an Exotoxin A specific single-stranded DNA (ssDNA) molecular recognition element (MRE). Its emphasis is on increasing stringency in directing binding toward free target of interest and at the same time decreasing binding toward negative targets. A ssDNA MRE with specificity and affinity was identified after fourteen rounds of Decoy-SELEX. Utilizing surface plasmon resonance measurements, the determined equilibrium dissociation constant (Kd) of the MRE is between 4.2 µM and 4.5 µM, and is highly selective for Exotoxin A over negative targets. A ssDNA MRE modified sandwich enzyme-linked immunosorbent assay (ELISA) has been developed and achieved sensitive detection of Exotoxin A at nanomolar concentrations in human serum. This study has demonstrated the proof-of-principle of using a ssDNA MRE as a clinical diagnostic tool.
    • Journal Subset:
      Biomedical; Peer Reviewed; USA
    • ISSN:
      2314-6133
    • MEDLINE Info:
      NLM UID: 101600173
    • Publication Date:
      20170724
    • Publication Date:
      20190517
    • DOI:
      http://dx.doi.org/10.1155/2015/417641
    • Accession Number:
      113630029
  • Citations
    • ABNT:
      HONG, K. L. et al. Selection of Single-Stranded DNA Molecular Recognition Elements against Exotoxin A Using a Novel Decoy-SELEX Method and Sensitive Detection of Exotoxin A in Human Serum. BioMed Research International, [s. l.], v. 2015, p. 1–12, 2015. DOI 10.1155/2015/417641. Disponível em: http://search.ebscohost.com/login.aspx?direct=true&site=eds-live&db=rzh&AN=113630029. Acesso em: 2 jul. 2020.
    • AMA:
      Hong KL, Yancey K, Battistella L, et al. Selection of Single-Stranded DNA Molecular Recognition Elements against Exotoxin A Using a Novel Decoy-SELEX Method and Sensitive Detection of Exotoxin A in Human Serum. BioMed Research International. 2015;2015:1-12. doi:10.1155/2015/417641.
    • AMA11:
      Hong KL, Yancey K, Battistella L, et al. Selection of Single-Stranded DNA Molecular Recognition Elements against Exotoxin A Using a Novel Decoy-SELEX Method and Sensitive Detection of Exotoxin A in Human Serum. BioMed Research International. 2015;2015:1-12. doi:10.1155/2015/417641
    • APA:
      Hong, K. L., Yancey, K., Battistella, L., Williams, R. M., Hickey, K. M., Bostick, C. D., Gannett, P. M., & Sooter, L. J. (2015). Selection of Single-Stranded DNA Molecular Recognition Elements against Exotoxin A Using a Novel Decoy-SELEX Method and Sensitive Detection of Exotoxin A in Human Serum. BioMed Research International, 2015, 1–12. https://doi.org/10.1155/2015/417641
    • Chicago/Turabian: Author-Date:
      Hong, Ka Lok, Kailey Yancey, Luisa Battistella, Ryan M. Williams, Katherine M. Hickey, Chris D. Bostick, Peter M. Gannett, and Letha J. Sooter. 2015. “Selection of Single-Stranded DNA Molecular Recognition Elements against Exotoxin A Using a Novel Decoy-SELEX Method and Sensitive Detection of Exotoxin A in Human Serum.” BioMed Research International 2015 (November): 1–12. doi:10.1155/2015/417641.
    • Harvard:
      Hong, K. L. et al. (2015) ‘Selection of Single-Stranded DNA Molecular Recognition Elements against Exotoxin A Using a Novel Decoy-SELEX Method and Sensitive Detection of Exotoxin A in Human Serum’, BioMed Research International, 2015, pp. 1–12. doi: 10.1155/2015/417641.
    • Harvard: Australian:
      Hong, KL, Yancey, K, Battistella, L, Williams, RM, Hickey, KM, Bostick, CD, Gannett, PM & Sooter, LJ 2015, ‘Selection of Single-Stranded DNA Molecular Recognition Elements against Exotoxin A Using a Novel Decoy-SELEX Method and Sensitive Detection of Exotoxin A in Human Serum’, BioMed Research International, vol. 2015, pp. 1–12, viewed 2 July 2020, .
    • MLA:
      Hong, Ka Lok, et al. “Selection of Single-Stranded DNA Molecular Recognition Elements against Exotoxin A Using a Novel Decoy-SELEX Method and Sensitive Detection of Exotoxin A in Human Serum.” BioMed Research International, vol. 2015, Nov. 2015, pp. 1–12. EBSCOhost, doi:10.1155/2015/417641.
    • Chicago/Turabian: Humanities:
      Hong, Ka Lok, Kailey Yancey, Luisa Battistella, Ryan M. Williams, Katherine M. Hickey, Chris D. Bostick, Peter M. Gannett, and Letha J. Sooter. “Selection of Single-Stranded DNA Molecular Recognition Elements against Exotoxin A Using a Novel Decoy-SELEX Method and Sensitive Detection of Exotoxin A in Human Serum.” BioMed Research International 2015 (November 9, 2015): 1–12. doi:10.1155/2015/417641.
    • Vancouver/ICMJE:
      Hong KL, Yancey K, Battistella L, Williams RM, Hickey KM, Bostick CD, et al. Selection of Single-Stranded DNA Molecular Recognition Elements against Exotoxin A Using a Novel Decoy-SELEX Method and Sensitive Detection of Exotoxin A in Human Serum. BioMed Research International [Internet]. 2015 Nov 9 [cited 2020 Jul 2];2015:1–12. Available from: http://search.ebscohost.com/login.aspx?direct=true&site=eds-live&db=rzh&AN=113630029