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Analytical and bioanalytical chemistry

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


PMID 27178553

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 0.400xa0pM, a limit of quantification of 1.70xa0pM, and an assay range spanning three orders of magnitude (1.70-900xa0pM) 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. Graphical abstract 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.