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Smartphone-based kanamycin sensing with ratiometric FRET
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 w...
Autores principales: | , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
The Royal Society of Chemistry
2019
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9060919/ https://www.ncbi.nlm.nih.gov/pubmed/35517283 http://dx.doi.org/10.1039/c8ra10035g |
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author | Umrao, Saurabh S, Anusha Jain, Vasundhara Chakraborty, Banani Roy, Rahul |
author_facet | Umrao, Saurabh S, Anusha Jain, Vasundhara Chakraborty, Banani Roy, Rahul |
author_sort | Umrao, Saurabh |
collection | PubMed |
description | 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. |
format | Online Article Text |
id | pubmed-9060919 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2019 |
publisher | The Royal Society of Chemistry |
record_format | MEDLINE/PubMed |
spelling | pubmed-90609192022-05-04 Smartphone-based kanamycin sensing with ratiometric FRET Umrao, Saurabh S, Anusha Jain, Vasundhara Chakraborty, Banani Roy, Rahul RSC Adv Chemistry 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. The Royal Society of Chemistry 2019-02-19 /pmc/articles/PMC9060919/ /pubmed/35517283 http://dx.doi.org/10.1039/c8ra10035g Text en This journal is © The Royal Society of Chemistry https://creativecommons.org/licenses/by-nc/3.0/ |
spellingShingle | Chemistry Umrao, Saurabh S, Anusha Jain, Vasundhara Chakraborty, Banani Roy, Rahul Smartphone-based kanamycin sensing with ratiometric FRET |
title | Smartphone-based kanamycin sensing with ratiometric FRET |
title_full | Smartphone-based kanamycin sensing with ratiometric FRET |
title_fullStr | Smartphone-based kanamycin sensing with ratiometric FRET |
title_full_unstemmed | Smartphone-based kanamycin sensing with ratiometric FRET |
title_short | Smartphone-based kanamycin sensing with ratiometric FRET |
title_sort | smartphone-based kanamycin sensing with ratiometric fret |
topic | Chemistry |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9060919/ https://www.ncbi.nlm.nih.gov/pubmed/35517283 http://dx.doi.org/10.1039/c8ra10035g |
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