Sensitive colorimetric sensor for point-of-care detection of acetylcholinesterase using cobalt oxyhydroxide nanoflakes

Abstract

Point-of-care monitoring of acetylcholinesterase (AChE) is of significant importance for pesticide poisoning and disease diagnosis because it plays a pivotal role in biological nerve conduction systems. Herein, we designed a colorimetric strategy for the facile and accurate detection of AChE based on tandem catalysis with a multi-enzyme system, which is constituted by cobalt oxyhydroxide nanoflakes (CoOOH NFs) and choline oxidase (CHO). In this sensor, AChE catalytically hydrolyzed acetylcholine (ACh) to produce choline, which was further efficiently oxidized by CHO to yield H₂O₂. CoOOH NFs, as a nanozyme, efficiently catalyzed 3,3′,5,5′-tetramethylbenzidine (TMB) into blue oxTMB with the help of H₂O₂, accompanied by an enhancement of absorbance intensity. The resulting intensity could be employed as the signal output of the CHO/CoOOH/ACh system in monitoring AChE. Under optimal conditions, the developed sensor possessed a sensitive response to AChE with a detection limit of 33 μU mL⁻¹. Interestingly, the proposed platform was applied to fabricate a paper-based sensor for rapidly recognizing AChE by direct observation with the naked eyes. Combined with a smartphone and ImageJ software, we further developed an image-processing algorithm for the quantitative detection of AChE with highly promising results, which validated the outstanding potential of on-site application in clinical diagnostics and pesticide poisoning.