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Aptasensor for ampicillin using gold nanoparticle based dual fluorescence–colorimetric methods

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Abstract

A gold nanoparticle based dual fluorescence–colorimetric method was developed as an aptasensor to detect ampicillin using its single-stranded DNA (ssDNA) aptamer, which was discovered by a magnetic bead-based SELEX technique. The selected aptamers, AMP4 (5′-CACGGCATGGTGGGCGTCGTG-3′), AMP17 (5′-GCGGGCGGTTGTATAGCGG-3′), and AMP18 (5′-TTAGTTGGGGTTCAGTTGG-3′), were confirmed to have high sensitivity and specificity to ampicillin (K d, AMP7 = 9.4 nM, AMP17 = 13.4 nM, and AMP18 = 9.8 nM, respectively). The 5′-fluorescein amidite (FAM)-modified aptamer was used as a dual probe for observing fluorescence differences and color changes simultaneously. The lower limits of detection for this dual method were a 2 ng/mL by fluorescence and a 10 ng/mL by colorimetry for ampicillin in the milk as well as in distilled water. Because these detection limits were below the maximum residue limit of ampicillin, this aptasensor was sensitive enough to detect antibiotics in food products, such as milk and animal tissues. In addition, this dual aptasensor will be a more accurate method for antibiotics in food products as it concurrently uses two detection methods: fluorescence and colorimetry.

The dual fluorescence-colorimetric method for the detection of ampicillin using the AMP17 ssDNA aptamer. The FAM-AMP17 aptamer is quenched by adsorption onto the surface of the AuNPs. If ampicillin and salt (NaCl) are added to the solution, fluorescence is recovered and the AuNPs aggregate. Conversely, the AuNP solution treated with only salt shows no obvious changes.

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Acknowledgements

This work was supported by the National Research Foundation of Korea (NRF-20100019914, 20110013431) and KOSEF through Center for Electro-Photo Behaviors in Advanced Molecular Systems (20110007166).

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Authors and Affiliations

  1. Department of Chemistry, Pohang University of Science and Technology, San 31, Hyoja-dong, Pohang, Gyungbuk, 790-784, South Korea

    Kyung-Mi Song, Euiyoung Jeong, Weejeong Jeon & Changill Ban

  2. Department of Mechanical Engineering, Department of Materials, University of California-Santa Barbara, Santa Barbara, CA, 93106, USA

    Minseon Cho

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  1. Kyung-Mi Song
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  2. Euiyoung Jeong
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  3. Weejeong Jeon
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  4. Minseon Cho
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Correspondence to Changill Ban.

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Song, KM., Jeong, E., Jeon, W. et al. Aptasensor for ampicillin using gold nanoparticle based dual fluorescence–colorimetric methods. Anal Bioanal Chem 402, 2153–2161 (2012). https://doi.org/10.1007/s00216-011-5662-3

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  • DOI: https://doi.org/10.1007/s00216-011-5662-3

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