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  • Title: A universal amplified strategy for aptasensors: enhancing sensitivity through allostery-triggered enzymatic recycling amplification.
    Author: Feng K, Kong R, Wang H, Zhang S, Qu F.
    Journal: Biosens Bioelectron; 2012; 38(1):121-5. PubMed ID: 22709934.
    Abstract:
    A universal amplified sensing strategy based on endonuclease was developed for designing fluorescence aptasensors. By employing hairpin-structured design for both recognition and reporter probes to decrease background signal, and a nicking endonuclease to perform target-triggered enzymatic recycling amplification, the proposed biosensor showed high sensitivity to target protein. To demonstrate the feasibility of the design, immunoglobulin E (IgE) was studied as a model target. Upon the addition of target protein, the specific formation of IgE/aptamer complex induced the releasing of the 37-mer fragment which partially hybridized with the molecular beacon (MB) probe. In the presence of endonuclease Nt.BbvCI, the MB was cleaved into two parts. Then, the released 37-mer fragment hybridized with another MB, and triggered the second cycle of cleavage, leading to an accumulation of fluorescence signals. Under the optimal conditions, a detection limit of 5 pM was obtained. The proposed sensing system was used for detection of IgE in complex biological samples with satisfactory results.
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