Signal-on electrogenerated chemiluminescence detection of gonyautoxin 1/4 based on proximity ligation-induced an electrode-bound pseudoknot DNA

Abstract

A “signal on” electrogenerated chemiluminescence (electrochemiluminescence, ECL) aptasensor based on proximity ligation-induced an electrode-bound pseudoknot DNA for sensitive detection of gonyautoxin 1/4 (GTX1/4) was developed on basis of the competitive type reaction mode. Aptamer was adopted as recognition element. Ru(bpy)32+ as ECL signal, was attached on the glassy carbon electrode (GCE) surface modified with nafion and gold nanoparticles (AuNPs) by electrostatic attraction to obtain the ECL platform. The pseudoknot DNA as capture probe, was immobilized onto the ECL platform via Au–S bond to obtain the ECL aptasensor. In the absence of GTX1/4, Y-shape proximate cooperative complex among aptamer, pseudoknot DNA and DNA1 was formed, drawing the ferrocene groups Fc, as ECL quencher) of both pseudoknot DNA and DNA1 near the electrode surface and resulting in low ECL signal. In the presence of GTX1/4, GTX1/4 competed with pseudoknot DNA and DNA1 for aptamer in homogeneous solution, preventing the formation of proximate cooperative complex and keeping the capture DNA in the pseudoknot conformation with Fc groups far away from the electrode surface, generating a high ECL signal. The recovery of ECL intensity increased with the GTX1/4 concentration and allowed the detection of GTX1/4 in the range of 0.01 ng/mL to 10 ng/mL with a detection of limit as low as 6.56 pg/mL. Additionally, the accuracy of this method was validated for analysis of spiked sea water samples with good recoveries, which indicates great potential in commercial application.