Journal
BIOSENSORS & BIOELECTRONICS
Volume 38, Issue 1, Pages 121-125Publisher
ELSEVIER ADVANCED TECHNOLOGY
DOI: 10.1016/j.bios.2012.05.008
Keywords
Aptasensors; Fluorescence; IgE; Enzymatic recycling amplification; Biological samples
Categories
Funding
- National Natural Science Foundation of China [21005047]
- Excellent Middle-age and Young Scientists Research Award Foundation of Shandong Province [BS2010SW012]
- Shandong provincial scientific instruments and equipment upgrading projects [2011SJGZ23]
- Colleges and universities of Shandong province science and technology plan projects [J12LD17]
- Guangdong Natural Science Foundation for Doctors [10451601501006188]
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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. (C) 2012 Elsevier B.V. All rights reserved.
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