GO-amplified fluorescence polarization assay for high-sensitivity detection of aflatoxin B1 with low dosage aptamer probe.

Abstract

Aflatoxin B1 (AFB1) is the most toxic mycotoxin of the aflatoxins (AFs) and shows carcinogenic, teratogenic and mutagenic effects in humans and animals. AFB1 is widely seen in cereal products such as rice and wheat. This research proposed a low-cost, high-sensitivity fluorescence polarization (FP) assay for detection of AFB1 using aptamer biosensors based on graphene oxide (GO). The aptamers labelled with fluorescein amidite (FAM) were adsorbed on the surface of GO through π-π stacking and electrostatic interaction, thus forming aptamer/GO macromolecular complexes. Under these conditions, the local rotation of fluorophores was limited and the system had a high FP value. When there was AFB1 in the system, aptamers were dissociated from the GO surface and combined with AFB1 owing to their specificity to form aptamer/AFB1 complexes. As a result, large changes were observed in the molecular weights of aptamers before, and after, the combination, therefore leading to the apparent changes in FP value. The results showed that when only 10nM of aptamer was used, the changes in FP and the AFB1 concentration had a favourable linear relationship within 0.05 to 5nM of AFB1, and the lowest detection limit (LOD) was 0.05nM. In addition, the recoveries of rice sample extract ranged from 89.2% to 112%. The method is simple, highly sensitive, cost-efficient and shows potential application prospects.