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Improving impedimetric nucleic acid detection by using enzyme-decorated liposomes and nanostructured screen-printed electrodes

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Abstract

Sensitive impedimetric detection of miR-222, a miRNA sequence found in many lung tumors, was investigated by using gold-nanostructured disposable carbon electrodes and enzyme-decorated liposomes. The proposed method was based on the immobilization of thiolated DNA capture probes onto gold-nanostructured carbon surfaces. Afterwards, the capture probes were allowed to hybridize to the target miRNAs. Finally, enzyme-decorated liposomes were used as labels to amplify the miRNA sensing, by their association with the probe–miRNA hybrids generated on the nanostructured transducer. By using this amplification route a limit of detection of 0.400 pM, a limit of quantification of 1.70 pM, and an assay range spanning three orders of magnitude (1.70–900 pM) were obtained (RSD % = 13). This limit of quantification was 20 times lower than that obtained using a simple enzyme conjugate for the detection. A comparison was also made with gold screen-printed transducers. In this case, a limit of quantification approximately 70 times lower was found by using the nanostructured transducers. Application of the optimized assay in serum samples was also demonstrated.

Alkaline Phosphatase-decorated liposomes and Au nanostructured screen-printed electrodes have been used for the impedimetric detection of miRNAs, via the bio-catalyzed precipitation of an insulating product onto the electrode surface.

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Acknowledgments

This work was supported by Ministero dell’Istruzione, dell’Università e della Ricerca (MIUR) in the framework of Progetti di ricerca di interesse nazionale (PRIN) 2012 (grant no. 20128ZZS2H) and Ente Cassa di Risparmio di Firenze project ID PED 8780 2014.0757A2202.0734.

EF and DB acknowledge CSGI (Consorzio per lo Sviluppo dei Sistemi a Grande Interfase) for partial financial support.

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Authors and Affiliations

  1. Dipartimento di Chimica, Università degli Studi di Firenze, Via della Lastruccia 3, Sesto Fiorentino, 50019, Firenze, Italy

    Diego Voccia, Francesca Bettazzi, Emiliano Fratini, Debora Berti & Ilaria Palchetti

  2. CSGI, University of Florence, Via della Lastruccia 3, Sesto Fiorentino, 50019, Firenze, Italy

    Emiliano Fratini & Debora Berti

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  1. Diego Voccia
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Correspondence to Ilaria Palchetti.

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Published in the topical collection Chemical Sensing Systems with guest editors Maria Careri, Marco Giannetto, and Renato Seeber.

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Voccia, D., Bettazzi, F., Fratini, E. et al. Improving impedimetric nucleic acid detection by using enzyme-decorated liposomes and nanostructured screen-printed electrodes. Anal Bioanal Chem 408, 7271–7281 (2016). https://doi.org/10.1007/s00216-016-9593-x

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  • DOI: https://doi.org/10.1007/s00216-016-9593-x

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