Fe3O4@polydopamine and Exo III-assisted homogeneous biorecognition reaction for convenient and ultrasensitive detection of kanamycin antibiotic.

Herein, we report a Fe3O4@polydopamine (PDA) nanocomposite and exonuclease III (Exo III)-assisted homogeneous fluorescence biosensing method for ultrasensitive detection of kanamycin (Kana) antibiotic. A hairpin DNA containing the Kana-aptamer sequence (HP) was first designed for the highly specific biorecognition of the target analyte. Because of the aptamer biorecognition-induced structural change of HP and the highly effective catalyzed reaction of Exo III, a large amount of fluorophore labels were released from the designed fluorescence DNA probe. During the homogeneous reaction process, the Exo III-assisted dual recycling significantly amplified the fluorescence signal output. Moreover, the excessive probes were easily adsorbed and separated by the Fe3O4@PDA nanocomposite, which decreased the background signal and increased the signal-to-noise ratio. These strategies result in the excellent analytical performance of the method, including a very low detection limit of 0.023 pg mL-1 and a very wide linear range of six orders of magnitude. In addition, this method has convenient operation, excellent selectivity, repeatability and satisfactory reliability, and does not involve the design and utilization of complicated DNA sequences. Thus, it exhibits a promising prospect for practical applications.

Exonuclease-assisted target recycling for ultrasensitive electrochemical detection of microRNA at vertically aligned carbon nanotubes.

As an important biomarker for early disease diagnosis, microRNA-21 (miRNA-21) has attracted considerable attention owing to its accurate detection. Herein we combine the one-step biorecognition reaction at a vertically aligned nanostructure-based biosensor with the T7 exonuclease (Exo)-assisted target recycling to develop a novel electrochemical bioassay method for miRNA-21 detection. The vertically aligned nanointerface is constructed through the covalent attachment of terminally carboxylated single-walled carbon nanotubes (SWCNTs) at an aryldiazonium salt-modified electrode, which enables the noncovalent adsorption of a ferrocene-labeled single-stranded signal DNA to obtain the biosensor. Upon its incubation with a target miRNA-21 solution, DNA/RNA hybridized duplexes will form and release from the electrode surface, leading to the corresponding electrochemical signal decrease of the biosensor. Moreover, this biorecognition reaction can also trigger the T7 Exo-assisted target recycling to achieve great signal amplification. Together with the highly efficient biorecognition and excellent electron transfer promotion at the vertically aligned SWCNTs, this biosensor exhibits a wide linear range varying from 0.01 to 100 pM and a low detection limit down to 3.5 fM. Considering its obvious performance superiority and convenient manipulations, this vertically aligned SWCNT-based electrochemical biosensing method has extensive potential for practical applications.