化学
核酸外切酶 III
检出限
DNA
杂交探针
组合化学
胶体金
DNA–DNA杂交
核酸外切酶
适体
A-DNA
生物物理学
分子生物学
纳米颗粒
纳米技术
色谱法
生物化学
DNA聚合酶
基因
材料科学
生物
大肠杆菌
作者
Ping Wang,Huikai Ma,Yanfeng Zhu,Wei Feng,Mengyao Su,Sanqiang Li,Hongju Mao
标识
DOI:10.1016/j.jelechem.2022.116167
摘要
In this work, a novel ratiometric electrochemical aptasensor was designed on the basis of dual-amplification mechanism by using exonuclease I (Exo I)-assisted target recycling and hybridization chain reaction (HCR). The thiol-modified capture DNA (Cp-DNA) was fixed on the surface of the gold electrode (AuE) modified with gold nanoparticles (AuNPs) through Au-S bonds. The ferrocene (Fc)-labeled DNA (Fc-DNA) hybridized with Cp-DNA to form Fc-DNA/Cp-DNA duplex. The presence of carcinoembryonic antigen (CEA) led to the dissociation of Fc-DNA from Fc-DNA/Cp-DNA duplex and initiated Exo Ⅰ-assisted target recycling to digest Fc-DNA, so numerous Fc left away from the electrode surface and resulting in the “signal off” of Fc. Moreover, Cp-DNA was exposed to bind trigger DNA and initiate HCR with the presence of H1 and H2 probes, resulting in the intercalating of numerous methylene blue (MB) molecules and the “signal-on” of MB. This dual signal strategy significantly amplified the decrease of Fc signal and the increase of MB signal, achieving a linear range of 1 pg mL−1 to 100 ng mL−1 with a detection limit of 0.479 pg mL−1 (S/N = 3). Besides, the developed aptasensor exhibited high specificity, good stability and reproducibility. More importantly, the aptasensor had been applied in detection of CEA in diluted human serum and displayed satisfactory anti-interference property. By changing the corresponding aptamer sequences, the designed strategy can be employed to detect other targets, which holds promising application in clinical diagnosis.
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