A sensitive colorimetric aptasensor for chloramphenicol detection in fish and pork based on the amplification of a nano-peroxidase-polymer†
Abstract
A novel colorimetric aptasensor was developed for sensitive and selective determination of chloramphenicol (CAP) based on gold nanoparticles (AuNPs) labeled with Power Vision (PV) and magnetic separation. PV, with a high enzyme-to-antibody ratio, is composed of a compact enzyme-linker antibody conjunction. In this assay, an aptamer of CAP was immobilized on Fe3O4@Au magnetic nanoparticles as a capture probe (AuMNPs–Apt) to concentrate target CAP. The complementary DNA (cDNA) and PV were both labeled on AuNPs to form a nano-peroxidase polymer as a signal tag (cDNA–AuNPs–PV). And the special tags could hybridize with the aptamer and cDNA to form AuMNP–Apt/cDNA–AuNP–PV conjugates. In the presence of CAP, the aptamer preferentially bound to CAP and caused the dissociation of some cDNA–AuNPs–PV on the conjugates with magnetic separation. PV, carried on signal tags, could greatly catalyze 3,3′,5,5′-tetramethylbenzidine (TMB) leading to color development, which could be quantified by Ultraviolet-visible (UV-vis) spectroscopy. A linear response to CAP concentration in the range of 0.05–200 ng mL−1 was obtained by this proposed method, with a low detection limit down to 0.02 ng mL−1. Besides, this assay was successfully employed to analyze CAP in fish and pork samples, whose results were consistent with those of the conventional enzyme-linked immunosorbent assay (ELISA) method.