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Sensitivity and Specificity of Metal Surface-Immobilized “Molecular Beacon” Biosensors
Abstract
The separate developments of microarray patterning of DNA oligonucleotides, and of DNA hairpins as sensitive probes for oligonucleotide identification in solution, have had a tremendous impact on basic biological research and clinical applications. We have combined these two approaches to develop arrayable and label-free biological sensors based on fluorescence unquenching of DNA hairpins immobilized on metal surfaces. The thermodynamic and kinetic response of these sensors, and the factors important in hybridization efficiency, were investigated. Hybridization efficiency was found to be sensitive to hairpin secondary structure, as well as to the surface distribution of DNA hairpins on the substrate. The identity of the bases used in the hairpin stem as well as the overall loop length significantly affected sensitivity and selectivity. Surface-immobilized hairpins discriminated between two sequences with a single base-pair mismatch with high sensitivity (over an order of magnitude difference in signal) under identical assay conditions (no change in stringency). This represents a significant improvement over other microarray-based techniques.
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Mathews, D. H., Sabina, J., Zuker, M., & Turner, D. H. (1999) · Journal of Molecular Biology · doi:10.1006/jmbi.1999.2700 · Cited by 3,795
Cited by
Borisov, S. M., & Wolfbeis, O. S. (2008) · Chemical Reviews · doi:10.1021/cr068105t · Cited by 1,018
Biosensor technology: recent advances in threat agent detection and medicine
Kirsch, J., Siltanen, C., Zhou, Q., Revzin, A., & Simonian, A. (2013) · Chemical Society Reviews · doi:10.1039/c3cs60141b · Cited by 462
Metadata
- Authors
- Du, H., Strohsahl, C. M., Camera, J., Miller, B. L., & Krauss, T. D.
- Venue
- Journal of the American Chemical Society
- Publisher
- American Chemical Society (ACS)
- Published
- 2005-05-05
- Volume
- 127
- Issue
- 21
- Pages
- 7932-7940
- ISSN
- 0002-7863 (ISSN-L)
- 1520-5126
- OpenAlex
- W2017359768
- Cited by
- 213
- References
- 1
- FWCI
- 5.19
- Type
- journal article
- Language
- en
- Open access
- closed
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- Du, H., Strohsahl, C. M., Camera, J., Miller, B. L., & Krauss, T. D. (2005). Sensitivity and Specificity of Metal Surface-Immobilized “Molecular Beacon” Biosensors. Journal of the American Chemical Society. https://doi.org/10.1021/ja042482a
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