In vitro selection of single-stranded DNA molecular recognition elements against S. aureus alpha toxin and sensitive detection in human serum Journal Article
| Authors: | Hong, K. L.; Battistella, L.; Salva, A. D.; Williams, R. M.; Sooter, L. J. |
| Article 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. |
| Keywords: | controlled study; nonhuman; sensitivity and specificity; binding affinity; polymerase chain reaction; bacteria (microorganisms); enzyme linked immunosorbent assay; molecular cloning; molecular recognition; dna sequence; bovine serum albumin; single stranded dna; staphylococcus aureus; exonuclease; elisa; surface plasmon resonance; aptamer; in vitro selection; pseudomonas exotoxin; dissociation constant; limit of detection; human; article; clostridium difficile toxin b; alpha toxin; molecular recognition element (mre); selex; ssdna; cholera toxin; staphylococcus alpha toxin |
| Journal Title: | International Journal of Molecular Sciences |
| Volume: | 16 |
| Issue: | 2 |
| ISSN: | 1422-0067 |
| Publisher: | MDPI |
| Date Published: | 2015-02-01 |
| Start Page: | 2794 |
| End Page: | 2809 |
| Language: | English |
| DOI: | 10.3390/ijms16022794 |
| PROVIDER: | scopus |
| PUBMED: | 25633102 |
| PMCID: | PMC4346865 |
| DOI/URL: | |
| Notes: | Export Date: 2 March 2015 -- Source: Scopus |
