Densitometry determination of dioxins using gold nanoparticlemodified dioxin response element probes

This study aims to develop a rapid and sensitive densitometry bioassay of dioxins. It is known that dioxin ligand can bind heterodimeric aryl hydrocarbon receptor (AhR) and trigger the formation of the complex of dioxin-AhR, AhR nuclear translocator (Arnt), and dioxin-responsive element (DRE). We have established a rapid and sensitive method using nanoparticle-modified DRE probes based on dioxin-dependent binding of DRE to AhR complex. Dioxins can trigger the binding of nanoparticle-modified DRE probe to AhR complex. The nanoparticle-modified DRE probes in the dioxin-AhR-Arnt-DRE complexes immobilized on the microplates were detected by measuring optical density after silver enhancement. Results showed that as little as 0.01 pM of TCDD could be detected. Dioxins could be quantified accurately with the linear range from 1 nM to 0.01 pM. The coefficient of variation was 5≈8% in this bioassay. The optical density of nanoparticle-based silver enhancement was produced in a dose- and time- dependent manner. The dioxin concentrations in fly ash samples determined by this bioassay were comparable with that by the standard HRGC/HRMS. Thus this bioassay is a sensitive and convenient cell-free bioassay of dioxin that is suitable for screening of dioxins in environmental or food matrices.