Visual Detection of Chlorpyrifos by DNA Hydrogel-Based Self-Actuated Capillary Aptasensor Using Nicking Enzyme-Mediated Amplification

The abuse of chlorpyrifos (CPF), an organophosphorus pesticide, poses significant health risks to humans. Therefore, rapid and accurate detection of residual CPF is crucial to human health due to its high risk in trace amounts. Herein, we developed a simple aptasensor that combines a DNA hydrogel-based self-driven capillary with nicking enzyme-mediated amplification (NEMA), in which the NEMA is triggered through the interaction of the aptamer with CPF, and then amplified to produce a large number of single-stranded DNA that can destroy the three-dimensional structure of the DNA hydrogel. Due to the different degrees of collapse of the hydrogel membrane structure, different amounts of liquid are adsorbed into the capillary under the action of surface tension, thus realizing the naked eye detection of CPF. Under optimal conditions, the DNA hydrogel-based self-actuated capillary aptasensor can sensitively detect chlorpyrifos in the concentration range of 1 ng/L to 1 mg/L, with a detection limit of 1.73 pg/L. The advantages of the aptasensor are simple conditions, high sensitivity, and a large detection concentration range, and only a thermostat and simple operation are needed to achieve its excellent analytical performance. In addition, the developed self-actuated capillary aptasensor was successfully applied for the determination of CPF in apple, grape, cabbage, and peanut kernel.