应用电化学还原法将固定在玻碳电极表面的氧化石墨还原为石墨烯,然后利用偶联活化剂将经过氨基修饰的急性早幼粒细胞白血病(APL)PML/RARα融合基因序列探针固定到石墨烯修饰电极表面,以亚甲基蓝(MB)为电化学杂交指示剂,并由差分脉冲伏安法检测人工合成APL的PML/RARα融合基因.结果表明,石墨烯对MB的检测信号起到了很好的增敏作用,杂交前后MB还原峰电流差值与靶标链DNA浓度在5×10-10~2.5×10-9 mol/L范围内呈线性关系,检出限为8×10-11 mol/L.该方法简单、特异性好,有望用于实际样品的检测.
A graphene modified glassy carbon electrode(GCE) was prepared by electrochemical reduction of graphite oxide fixed on the surface of GCE.Then,the amino-group modified probe of acute promyelocytic leukemia(APL) PML/RARα fusion gene fragment was immobilized onto surface of the graphene modified GCE by coupling activating agent.With methylene blue(MB) as a novel electrochemical indicator,artificial APL PML/RARα fusion gene fragment was measured by differential pulse voltammetry(DPV).The results indicated that graphene nanomaterials showed good sensitivity to the signal of MB.The relationship between the reduction peak current of MB and the concentration of complementary strand was linear in the range of 5×10-10~2.5×10-9 mol/L.The detection limit had been estimated to be 8×10-11 mol/L.The method is simple and has good specificity and high sensitivity.This approach is promising to the detection of APL in the real sample for medical diagnostics.