DNA序列氧化损伤的电致化学发光定量检测

8-OHdG is the oxidative product of guanine in DNA sequence, which has been used as one of the main biomarkers for DNA oxidative damage, and also for diagnosis of oxidative stress related diseases. Current detection methods are mainly focused on the measurement of 8-OHdG existed in body fluid or tissue, which reflect only those 8-OHdG cleaved form DNA sequence. While, how many G has been oxidized in DNA sequence? what is the ratio between 8-OHdG and G in DNA sequence? That is DNA sequence oxidation damage rate here, and this value represents the real situation of DNA oxidative damage. Unfortunately, current detection methods can not be employed to fulfill this. To solve this problem, in the proposal, a series of osmium/ruthenium trisbipyridyl complexes with flexible saturated carbon chain are designed to covalently link with carbon nanotube (CNT) to give CNT chemically modified ECL labels. After the CNT modified ECL labels have been coated on the surface of the working electrode, ECL can be measured for the corresponding DNA sequence, the ployGC chain and the standard sample of 8-OHdG, respectively. Then the ratio between 8-OHdG and G in DNA sequence can be calculated based on the calibration curve and the ECL acquired from both osmium and ruthenium complexes modified CNT labels. An ECL method with independent intellectual property rights will be built up and employed for evaluation of DNA oxidative damage rate, which can be further utilized to provide reliable quantitative data for study of DNA oxidative damage and diagnosis of oxidative stress related diseases.

8-羟基脱氧鸟苷(8-OHdG)是DNA碱基鸟嘌呤G的氧化产物,也是癌变早期DNA氧化损伤的生物标示物，可用于氧化应激相关疾病的诊断。目前的检测方法更多侧重于对体液、相关组织器官中8-OHdG具体数量的测定，仅代表从DNA序列上切除下来的8-OHdG。而DNA序列中到底有多少G已被氧化?或者序列中8-OHdG与G的比例是多少？这一数值更能反映DNA序列氧化损伤的真实状况，也就是DNA序列氧化损伤率。现有方法不能满足其检测要求。本申请分别将联吡啶锇/钌电致化学发光(ECL)标示物与碳纳米管共价接枝，涂敷到工作电极表面。分别对DNA样本序列进行ECL检测，参照polyGC、8-OHdG标准品的ECL工作曲线,计算得到DNA序列的氧化损伤率，建立评价机体DNA氧化损伤真实程度的有效方法，为DNA氧化损伤的研究及氧化应激相关疾病的诊断提供可靠的量化指标。可获得自主知识产权,具有理论和实用价值。

8-羟基脱氧鸟苷(8-OHdG)是DNA碱基鸟嘌呤G的氧化产物,也是癌变早期DNA氧化损伤的生物标示物，可用于氧化应激相关疾病的诊断。目前的检测方法更多侧重于对体液、相关组织器官中8-OHdG具体数量的测定，仅代表从DNA序列上切除下来的8-OHdG。而DNA序列中到底有多少G已被氧化?或者序列中8-OHdG与G的比例是多少？这一数值更能反映DNA序列氧化损伤的真实状况，也就是DNA序列氧化损伤率。现有方法不能满足其检测要求。本申请分别将联吡啶锇/钌电致化学发光(ECL)标示物与碳纳米管共价接枝，涂敷到工作电极表面。分别对DNA样本序列进行ECL检测，参照polyGC、8-OHdG标准品的ECL工作曲线,计算得到DNA序列的氧化损伤率，建立评价机体DNA氧化损伤真实程度的有效方法，为DNA氧化损伤的研究及氧化应激相关疾病的诊断提供可靠的量化指标。构建了ECL检测盐酸氯丙嗪、喹诺酮抗生素、三聚氰胺、孔雀石绿、Cu2+、Hg2+、Zn2+等金属离子，以及荧光检测次氯酸/硫化氢可逆氧化还原、除草剂百草枯等方法。发表SCI论文9篇，获授权中国发明专利2件,具有理论和实用价值。