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应用恒电位在金基底表面电化学沉积纳米金,通过Au—S键将巯基修饰DNA探针固定在纳米金表面,与互补靶序列杂交,构建计时库仑电化学DNA传感器,并检测急性早幼粒细胞白血病(APL)PML/RARα融合基因.采用扫描电子显微术(SEM)与电化学交流阻抗技术(EIS)观察纳米金和表征DNA传感器的构筑过程.以氯化六氨合钌([Ru(NH3)6]Cl3,RuHex)作电化学杂交指示剂,由计时库仑法检测人工合成APL的PML/RARα融合基因.结果表明,纳米金能放大RuHex检测信号,杂交前后电量差值(ΔQ)与靶标链DNA浓度的对数(lgC)值在1.0×10-13～1.0×10-9mol.L-1范围内呈线性关系,检出下限3.7×10-14mol.L-1(S/N=3).该法操作简便、特异性好,有望用于实际样品的检测.
Gold nanoparticles were electrochemically deposited on the gold substrate by potentiostatic potentials. The thiol-modified DNA probes were immobilized on gold nanoparticles by Au-S bonds and hybridized with complementary target sequences to construct chrono-coulomb electrochemical DNA sensors. The detection of acute promyelocytic (APL) PML / RARα fusion gene.We used scanning electron microscopy (SEM) and electrochemical impedance spectroscopy (EIS) to observe the gold nanoparticles and characterize the construction of DNA sensors.Using ruthenium hexachloride ([Ru (NH3) 6] Cl3, RuHex as electrochemical hybridization indicator, the chimeric coulometry was used to detect the PML / RARα fusion gene of synthetic APL.The results showed that nano gold could amplify the signal of RuHex, (LgC) in the range of 1.0 × 10-13 ~ 1.0 × 10-9mol.L-1, the detection limit was 3.7 × 10-14mol.L-1 (S / N = 3). The method is simple, good specificity, is expected to be used for the actual sample testing.