Journal
BIOSENSORS & BIOELECTRONICS
Volume 29, Issue 1, Pages 66-75Publisher
ELSEVIER ADVANCED TECHNOLOGY
DOI: 10.1016/j.bios.2011.07.068
Keywords
Aptamer; Biofunctional probe; Label-free; Amplification detection
Categories
Funding
- National Natural Science Foundation of China [90817101, 20905022, 20775023]
- Guangdong Natural Science Foundation [10451601501006188]
- Huizhou University Foundation [C5090202]
- 973 National Basic Research Program of China [2007CB310500]
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Oligonucleotide-based detection schemes that avoid chemical modification possess significant advantages, including simplified design, intrinsic affinity for targets, low cost and ease to extend applications. In this contribution, we developed a label-free self-locked bifunctional oligonucleotide probe (signaling probe) for the detection of different disease markers in parallel. Two signal enhancement techniques based on isothermal circular strand-displacement polymerization reaction, cyclical nucleic acid strand-displacement polymerization (CNDP) and cyclical common (nonnucleic acid) target-displacement polymerization (CCDP), were employed to implement the amplification assay for p53 gene and PDGF-BB, respectively. The attractive assay properties confirmed the effectiveness of isothermal polymerization in common biosensing systems without evolving any chemical modification: PDGF could be detected down to 0.87 ng/mL, and a dynamic response range of 8-5000 ng/mL was achieved; The capability to screen the p53 gene was also considerably improved, including the detection limit, sensitivity, dynamic range and so on. Moreover, because no any chemical modification of the signaling probe was acquired and different targets were separately detected in homogeneous solution. This interrogating platform exhibits the design flexibility, convenience, simplicity and cost-effectiveness. The success achieved here is expected to serve as a significant step toward the development of robust label-free oligonucleotide probes in biomarker profiling and disease diagnostics. (C) 2011 Elsevier B.V. All rights reserved.
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