Umrao, S and Anusha, S and Jain, V and Chakraborty, B and Roy, R (2019) Smartphone-based kanamycin sensing with ratiometric FRET. In: RSC Advances, 9 (11). pp. 6143-6151.
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Abstract
Smartphone-based fluorescence detection is a promising avenue for biosensing that can aid on-site analysis. However, quantitative detection with fluorescence in the field has been limited due to challenges with robust excitation and calibration requirements. Here, we show that ratiometric analysis with Förster resonance energy transfer (FRET) between dye pairs on DNA aptamers can enable rapid and sensitive kanamycin detection. Since our detection scheme relies on ligand binding-induced changes in the aptamer tertiary structure, it is limited only by the kinetics of ligand binding to the aptamer. Our FRET-based kanamycin binding aptamer (KBA) sensor displays two linear ranges of 0.05-5 nM (detection limit of 0.18 nM) and 50-900 nM of kanamycin. The aptamer displays high specificity even in the presence of the 'natural' background from milk. By immobilizing the aptamer in the flow cell, our KBA sensor design is also suitable for repeated kanamycin detection. Finally, we show that the ratiometric FRET-based analysis can be implemented on a cheap custom-built smartphone setup. This smartphone-based FRET aptamer scheme detects kanamycin in a linear range of 50-500 nM with a limit of detection (LOD) of 28 nM. © 2019 The Royal Society of Chemistry.
| Item Type: | Journal Article |
|---|---|
| Publication: | RSC Advances |
| Publisher: | RSC |
| Additional Information: | Copyright for this article belongs to RSC. |
| Keywords: | Binding energy; Energy transfer; Forster resonance energy transfer; Ligands, Detection limits; Fluorescence detection; High specificity; Limit of detection; Quantitative detection; Ratiometric analysis; Resonance energy transfer; Tertiary structures, Smartphones |
| Department/Centre: | Division of Biological Sciences > Molecular Biophysics Unit Division of Interdisciplinary Sciences > Centre for Biosystems Science and Engineering Division of Mechanical Sciences > Chemical Engineering |
| Date Deposited: | 20 Mar 2019 05:54 |
| Last Modified: | 20 Mar 2019 05:54 |
| URI: | http://eprints.iisc.ac.in/id/eprint/62024 |
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