Alcohol dehydrogenase coexisted solid-state electrochemiluminescence biosensor for detection of p53 gene  

乙醇脱氢酶共存固相电致化学发光生物传感器对p53基因的检测(英文)

作　　者：王晓英[1] 王晓宁[2] 张相依[1] 陈奋天[1] 朱柯蕙[1] 杨立刚[1] 唐萌[1] 

机构地区：[1]东南大学环境医学工程教育部重点实验室,南京210009 [2]西安交通大学第一附属医院血液科,西安710061

出　　处：《Journal of Southeast University(English Edition)》2013年第2期145-151,共7页东南大学学报（英文版）

基　　金：The National Basic Research Program of China(973 Program)(No.2010CB732404,2011CB933404);the National Natural Science Foundation of China(No.81172697,81170492,81001244);the Specialized Research Fund for the Doctoral Program of Higher Education(No.20110092120055);the Foundation of the State Key Laboratory of Bioelectronics of Southeast University

摘　　要：An alcohol dehydrogenase （ADH）-coexisted solidstate electrochemiluminescence （ECL） biosensor for sensitive detection of the p53 gene was developed. The electrode modified by multiwalled carbon nanotubes, Ru（bpy）]2＋3 and polypyrrole （ MWNTs-Ru （bpy） ]2＋3 -PPy ） was prepared to adsorb the ssDNA by electrostatic interactions. Then, the ssDNA recognized the gold nanoparticles （AuNPs）-labeled p53 gene and produced the AuNPs-dsDNA electrode with the AuNPs layer. The AuNPs layer adsorbed the ADH molecules for producing the ECL signal. Thus, the biosensor was based on coupling enzyme substrate reaction with solid-state ECL detection, and it displayed good sensitivity and specificity. The detection limit of the wild type p53 sequence （wtp53） is as low as 0. 1 pmol/L and the discrimination is up to 57. 1% between the wtp53 and the muted type p53 sequence （mtp53）. The amenability of this method to the analyses of p53 from normal and cancer cell lysates is demonstrated. The signal of wtp53 in the MGC-803 gastric cancer cell lysates turns out to be about 61.8% that of the wtp53 in the GES-1 normal gastric mucosal cell lysates, and the concentration of the wtp53 is found to decrease about 59 times. The method is highly complementary to enzyme-linked immunosorbent assay （ELISA）, and it holds promise for the diagnosis and management of cancer.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.

关 键 词：MWNTs-Ru （ bpy ）2＋3 composite solid-stateelectrochemiluminescence alcohol dehydrogenase wild typep53 sequence muted type p53 sequence cell lysates 

分 类 号：R113[医药卫生—公共卫生与预防医学]