乙醇脱氢酶共存固相电致化学发光传感器检测p53基因
首发时间:2012-06-01
摘要:通过羧基化多壁碳纳米管(COOH-MWNTs)将电致化学发光活性物2,2'-联吡啶钌固定,经吡咯膜(PPy)把碳纳米管-联吡啶钌混聚体与纳米金胶标记的p53分析物相连,纳米金胶进一步偶联乙醇脱氢酶(ADH),将乙醇脱氢酶参与的专一性的酶促反应与高灵敏的电致化学发光反应成功偶联,建立一种直接、准确的酶共存固相电致化学发光检测人类p53肿瘤抑制基因的方法。用此法检测目标p53野生型序列,最低检测限为0.1 pmol/L,在相同条件下,单碱基错配的p53野生型序列和p53突变型序列的区分度达57.1%,其特异性、低灵敏度的检测为癌症的早期诊断提供了可能。
关键词: 电致化学发光 碳纳米管-联吡啶钌混聚体 乙醇脱氢酶 p53野生型 p53突变型
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Alcohol dehydrogenase coexisted solid-state electrochemiluminescence biosensor for detection of p53 tumor suppressor gene
Abstract:An alcohol dehydrogenase (ADH)-coexisted solid-state electrochemiluminescence (ECL) biosensor for sensitive detection of p53 tumor suppressor gene is developed successfully. A composite of multiwalled carbon nanotubes and Ruthenium (II) tris-(bipyridine) was prepared and coated on an glassy carbon (GC) electrode through hydrophobic interaction, which was covered by polypyrrole (PPy) to immobilize ssDNA. Then, the ssDNA recognized the gold nanoparticles (AuNPs)-labeled p53 tumor suppressor gene, and produced AuNPs-dsDNA electrode with AuNPs layer. Such surface adsorbed the ADH molecules for producing ECL signal. Thus, such biosensor was based on coupling enzyme substrate reaction with solid-state ECL detection, and it displayed good sensitivity and specificity. The wild type p53 sequence (wtp53) as low as 0.1 pmol/L can be detected and the discrimination is up to 57.1% between the wtp53 and the muted type p53 sequence (mtp53). It holds promise for the early diagnosis of cancer development and monitoring of patient therapy.
Keywords: Electrochemiluminescence carbon nanotubes-Ruthenium (II) tris-(bipyridine) composite alcohol dehydrogenase wild type p53 sequence muted type p53 sequence
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