Abstract As a significant potential biomarker for early disease diagnostics, the detection of 5-hydroxymethylcytosine (5hmC) has garnered widespread attention. In this work, a simple and sensitive electrochemiluminescence (ECL) was developed for 5hmC detection. After substrate electrode preparation, anti-5hmC antibody was captured on electrode surface by the formation of amide bond. Then, through immunoreaction, 5hmC was specifically recognized and captured. Utilizing the crosslink effect of Zr4+ with phosphoric acid recognition, aminomethyl phosphate was modified on electrode to further capture PAMAM‑silver nanoclusters/Nitrogen-doped graphene nanocomposite (PAMAM-Ag-NG), where this nanocomposite was employed as ECL signal probe. The designed immunosensor showed a linear concentration range from 10 pM to 30 nM with a low detection limit of 2.47 pM (S/N = 3). This method presented good selectivity and reproducibility. To prove the applicability, this method was further used to study the influence of phytohormone and avian leukemia virus on 5hmC content in maize seedling leaves and chicken fibroblast cells, respectively.

中文翻译：

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基于PAMAM-纳米银-氮掺杂石墨烯纳米复合材料的5-羟甲基胞嘧啶检测电化学发光免疫传感器

摘要 作为早期疾病诊断的重要潜在生物标志物，5-羟甲基胞嘧啶（5hmC）的检测受到广泛关注。在这项工作中，开发了一种用于 5hmC 检测的简单而灵敏的电化学发光 (ECL)。基板电极制备后，抗 5hmC 抗体通过形成酰胺键被捕获在电极表面。然后，通过免疫反应，5hmC 被特异性识别和捕获。利用 Zr4+ 与磷酸识别的交联效应，在电极上修饰磷酸氨基甲酯以进一步捕获 PAMAM-银纳米团簇/氮掺杂石墨烯纳米复合材料 (PAMAM-Ag-NG)，该纳米复合材料用作 ECL 信号探针。设计的免疫传感器显示线性浓度范围从 10 pM 到 30 nM，低检测限为 2。47 pM (S/N = 3)。该方法具有良好的选择性和重现性。为证明该方法的适用性，进一步研究植物激素和禽白血病病毒对玉米幼苗叶片和鸡成纤维细胞中5hmC含量的影响。