Rapid electrochemical biosensor composed of DNA probe/iridium nanoparticle bilayer for Aphanizomenon flos-aquae detection in fresh water

Toxic cyanobacteria pose a serious threat to aquatic ecosystems and require adequate detection and control systems. Aphanizomenon flos-aquae is a harmful cyanobacterium that produces the toxicant saxitoxin. Therefore, it is necessary to detect the presence of A. flos-aquae in lakes and rivers. We proposed a rapid electrochemical biosensor composed of DNA primer/iridium nanoparticles (IrNP) bilyer for the detection of A. flos-aquae in freshwater. The extracted A. flos-aquae gene (rbcL-rbcX) is used as a target, and it was fixed to the electrode using a 5'-thiolated DNA primer (capture probe). Then, Avidin@IrNPs complex for amplification of electrical signals was bound to the target through a 3'-biotinylated DNA primer (detection probe). To rapidly detect the target, an alternating current electrothermal flow technique was introduced in the detection step, which could reduce the detection time to within 20 min. To confirm the biosensor fabrication, atomic force microscopy was used to investigate the surface morphology. To evaluate the biosensor performance, cyclic voltammetry and electrochemical impedance spectroscopy were used. The target gene was detected at a concentration of 9.99 pg/mL in tap water, and the detection range was 0.1 ng/mL to 10³ ng/mL with high selectivity. Based on the combined system, we employed A. flos-aquae in tap water. This rapid cyanobacteria detection system is a powerful tool for CyanoHABs in the field.