Ultrasensitive aflatoxin B1 assay based on FRET from aptamer labelled fluorescent polymer dots to silver nanoparticles labeled with complementary DNA

Abstract

The authors describe a new method for the selective detection of aflatoxin B1 (AFB1) by an off-on signaling procedure in a fluorescence resonance energy transfer (FRET)based nanobioprobe. An amino-modified aptamer against AFB1 was conjugated to fluorescent polymer dots, containing poly(9,9-dioctylfluorenyl-2,7-diyl)-co-(1,4-benzothiadiazole) as the fluorophore. Complementary DNA (cDNA) was conjugated to silver nanoparticles (cDNA-AgNPs) which act as FRET acceptors. Mixed in solution, in the absence of AFB1, the aptamer and its cDNA hybridize to form (aptamer-cDNA). This brings the polymer dots into close proximity of the AgNPs and result in FRET from the donor to the acceptor due to spectral overlap between the emission of the polymer dots and the absorption of the AgNPs. The fluorescence of the polymer dots probe is thereby switched off. However, in the presence of AFB1, the aptamer with high affinity for AFB1 will be released from the cDNA-AgNP aggregate, which results in recovery of fluorescence (Bswitch on" state). The yellow fluorescence of the polymer dots, best measured at 538 nm, increases linearly in the 5 pg.mL(-1) to 1.0 ng.mL(-1) AFB1 concentration range, with a 0.3 pg.mL(-1) detection limit. The assay was successfully applied to the detection of AFB1 in (spiked) wheat flour, and the results were found to be in satisfactory agreement with those obtained by an enzyme-linked immunosorbent assay.