DNAzyme-Fluorescence Resonance Energy Transfer (FRET) Based Rapid Detection of Lead Pollution in Food
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Abstract:
To achieve rapid detection of lead (Pb2+) pollution in food samples, here, a simple “Turn on” Pb2+ sensor based on DNAzyme and Fluorescence Resonance Energy Transfer (FRET) was established. The quencher BHQ1 worked as the FRET acceptor and quenched the fluorescence of EvaGreen. In the presence of Pb2+, the specific cleavage of the substrates weakened the reaction between EvaGreen and BHQ1, the quencher BHQ1 released as the substrate cleaved and was far away from the FRET donor EvaGreen, so the FRET effect was weakened, the fluorescence of the donor EvaGreen was retained. The weakening of FRET effectand the enhancement of EvaGreen fluorescence signal induced by Pb2+could be used for qualitative and quantitative analysis of Pb2+pollution. When the armlength of the DNAzyme was 15 nt~5 nt, the concentration of EvaGreen, Substrate (GR-B) and DNAzyme were 1×, 400 nmol/L and 100 nmol/L, respectively, the limit of detection and linear range were explored. The reaction could be finished within 3 minutes at room temperature and had a high specificity for Pb2+, which could distinguish different metal ions. And it has been successfully applied to the quantitative analysis of Pb2+ in fresh eggs and tap water samples, which showed a satisfactory recovery rate from 86.79 % to 113.32 %. In summary, the proposed “Turn on” sensor could be used for rapid one-tube test at room temperature, amplification was not required, which might provide some ideas for detecting Pb2+pollution in food.
