A novel fluorescent assay based on DNAzyme-assisted detection of prostate specific antigen for signal amplification

Prostate specific antigen (PSA) is one of the most common biomarkers for the management of prostate cancer. However, it still remains urgent to develop highly sensitive, cost-effective and selective strategies for PSA assay. In this paper, we developed a low-cost, highly sensitive and specific analytical strategy for the detection of PSA by using a fluorescence sensor based on Pb²⁺-dependent DNAzyme. We designed a DNA sequence called cmMB with a hairpin structure, containing PSA-specific aptamers and Pb²⁺-dependent DNAzyme chains. Also, a fluorophore-labelled DNA sequence called Sub-FAM, which contains a cleavage site of Pb²⁺-dependent DNAzyme and serves as substrate, is also designed for the signal generation. In the presence of PSA, interaction between aptamer and PSA blocks the hairpin structure of cmMB, resulting in the formation of Pb²⁺-dependent DNAzyme with Pb²⁺. Then, Pb²⁺-dependent DNAzyme can cleavage Sub-FAM and produce a high fluorescence. In the absence of PSA, since Sub-FAM remains to be ssDNA and can be absorbed by GO, only low fluorescence can be detected. Under optimal experimental conditions, a good linear relationship in the range of 1-100 pg mL^-1 was exhibited, with a limit of detection (LOD) of 0.76 pg mL^-1. In addition, the proposed method has potential value in the diagnosis and monitoring of prostate cancer because of its good selectivity and practical application in biological samples.