期刊
ANALYTICAL CHEMISTRY
卷 87, 期 20, 页码 10385-10391出版社
AMER CHEMICAL SOC
DOI: 10.1021/acs.analchem.5b02495
关键词
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资金
- NNSF of China [21575116, 51473136]
- Fundamental Research Funds for the Central Universities [XDJK2014C138]
Graphene quantum dots (GQDs) with an average diameter as small as 2.3 nm were synthesized to fabricate an electrochemiluminescence (ECL) biosensor based on T7 exonuclease-assisted cyclic amplification and three-dimensional (3D) DNA-mediated silver enhancement for microRNA (miRNA) analysis. Herein, to overcome the barrier in immobilizing GQDs, aminated 3,4,9,10-perylenetetracarboxylic acid (PTCA NH2) was introduced to load GQDs through pi-pi stacking (GQDs/PTCA NH2), realizing the solid-state GQDs application. Furthermore, Fe3O4 Au core shell nanocomposite (Au@Fe3O4) was adopted as a probe anchor to form a novel electrochemiluminescent signal tag of GQ_Ds/PTCA NH2/AupFe(3)O(4). The prepared ECL signal tag was decorated on the electrode surface, exhibiting excellent film-forming performance, good electronic conductivity, and favorable stability, all of which overcame the obstacle for applying GQDs in ECL biosensing and showed a satisfactory ECL response under the coreactant of S2O82- (peroxydisulfate). Afterward, hairpin probe modified on the electrode was opened by helper DNA, followed by assembling target to hybridize with the exposed stem of the helper DNA. Significantly, T7 exonuclease was employed to digest the DNA/RNA duplex and trigger the target recycling without asking for a specific recognition site in the target sequence, realizing a series of RNA/DNA detections by changing the sequence of the complementary DNA. At last, the ECL signal was further enhanced by silver nanoparticles (AgNPs)-based 3D DNA networks. After the two amplifications, the ECL signal of GQDs was extraordinarily increased and the prepared biosensor achieved a high sensitivity with the detection limit of 0.83 fM. The biosensor was also explored in real samples, and the result was in good accordance with the performance of quantitative real-time polymerase chain reaction (qRT-PCR). Considering the excellent sensitivity and applicability, we believe that the proposed biosensor is a potential candidate for nucleic acid biosensing.
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