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构建乙醇脱氢酶(ADH)共存固相电致化学发光(ECL)生物传感器,对p53基因进行检测.多壁碳纳米管、Ru(bpy)2+3及吡咯膜复合物(MWNTs-Ru(bpy)2+3-PPy)修饰电极可有效固载ssDNA,ssDNA与纳米金胶(AuNPs)标记的p53基因杂交,AuNPs进而吸附ADH.将ADH参与的专一性的酶促反应与高灵敏的固相ECL技术偶联,建立检测p53基因的方法.该方法对p53野生型序列(wtp53)的检测限为0.1pmol/L,对p53突变序列(mtp53)和wtp53的区分度达57.1%.应用该方法,MGC-803胃癌细胞溶解产物中的wtp53产生信号强度仅为正常GES-1胃粘膜细胞溶解产物中wtp53产生信号强度的61.8%,浓度降低约59倍.该方法与传统的ELISA法具有较好的互补性,其在癌症早期诊断、临床治疗过程监控等方面具有潜在的应用前景.
(ADH) coexisting solid-phase electrochemiluminescence (ECL) biosensors were constructed to detect p53 gene.Multi-walled carbon nanotubes, Ru (bpy) 2 +3 and MWNTs-Ru (bpy) 2 + 3-PPy) modified electrode can effectively immobilize ssDNA, ssDNA hybridize with AuNPs labeled p53 gene, and AuNPs adsorb ADH.The specific enzymatic reaction involved in ADH is associated with highly sensitive The method was applied to detect p53 gene by solid-phase ECL technique.The detection limit of p53 wild-type sequence (wtp53) was 0.1 pmol / L, and the discrimination between p53 mutant sequence (wtp53) and wtp53 was 57.1% In this method, the signal strength of wtp53 in MGC-803 gastric cancer cell lysate was only 61.8% and the concentration was reduced by about 59-fold compared with that of wtp53 in normal GES-1 gastric mucosal lysate.This method has the same Better complementarity, which has potential applications in the early diagnosis of cancer, the monitoring of clinical treatment process and so on.