Chronocoulometric aptamer based assay for staphylococcal enterotoxin B by target-triggered assembly of nanostructured dendritic nucleic acids on a gold electrode
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A rapid and ultrasensitive method is described for the detection of staphylococcal enterotoxin B (SEB). It is based on the formation of a dendritic DNA superstructure by integrating (a) target-induced triggering of DNA release with (b) signal amplification by a hybridization chain reaction. Partially complementary pairing of aptamer and trigger DNA forms a duplex structure. The capture DNA is then placed on the surface of a gold electrode through gold-thiol chemistry. In the presence of SEB, the aptamer-target conjugate is compelled to form. This causes the release of trigger DNA owing to a strong competition with SEB. The trigger DNA is subsequently hybridized with the partial complementary sequences of the capture DNA to trigger HCR with three auxiliary DNA sequances (referred to as H1, H2, H3). Finally, the dendritic DNA superstructure is bound to hexaammineruthenium(III) cation by electrostatic adsorption and assembled onto the modified gold electrode. This produces an amplified electrochemical signal that is measured by chronocoulometry. Under optimal conditions, the charge difference increases linearly with the logarithm of the SEB concentrations in the range from 5 pg·mL−1 to 100 ng·mL−1 with a detection limit as low as 3 pg·mL−1 (at S/N = 3).
KeywordsStaphylococcus aureus Dendritic nucleic acid nanostructure Hybridization chain reaction Electrochemical nanoprobe ELISA Contaminated food Toxic protein Ruthenium(III) hexammine Electrostatic adsorption
The project was supported by the National Key Research and Development Program of China (No. 2018YFC1602800), the National Natural Science Foundation of China (No. 21804071), Natural Science Foundation of Jiangsu Province of China (No. BK20180688), and the University Science Research Project of Jiangsu Province in China (16KJB550005).
Compliance with ethical standards
The author(s) declare that they have no competing interests.
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