Journal
RSC ADVANCES
Volume 5, Issue 80, Pages 65437-65443Publisher
ROYAL SOC CHEMISTRY
DOI: 10.1039/c5ra11271k
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Funding
- National Natural Science Foundation of China [21205040, 21335003, 21421004]
- Chinese Ministry of Education [313019]
- Shanghai Fund [12ZR1442700]
- Fundamental Research Funds for the Central Universities
- Hitachi, Ltd.
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Developing label-free molecular beacon (MB)-based methods for DNA detection has been of great significance in bioanalysis because of their simplicity, low cost, and specificity. In this work, we have developed a novel DNA-scaffolded silver nanocluster linear molecular beacon (AgNC-LMB)-based strategy for sequence-specific DNA detection via exonuclease III (Exo 110-assisted signal amplification. The proposed method involves two processes: target-mediated digestion by Exo III and the synthesis of AgNC-LMB as a switch for signal output. Upon hybridization of the rationally designed probe with target, Exo III, removes nucleotides from 3' terminus of the probe. The resultant fragment acts as a favorable template to form highly fluorescent DNA/AgNCs. The resulting fluorescence enhancement of the AgNCs provides a quantitative readout proportional to the target concentration in the range of 5 to 300 nM with an LOD of similar to 3.2 nM. This method is simple, cost-effective, highly selective, and free of modification or separation.
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