Photoelectrochemical sensor based on AuNPs@WO3@TpPa-1-COF for quantification of DNA methylation levels
Photoelectrochemical sensor based on AuNPs@WO3@TpPa-1-COF for quantification of DNA methylation levels
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DOI：
10.1007/s00604-024-06235-1
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发表时间：
2024-03-01
影响因子：
5.7
通讯作者：
Zheng，Junsong
中科院分区：
文献类型：
--
作者：
Liu，Huamin;Li，Chenghong;Zheng，Junsong
设计了一种用于超灵敏检测DNA甲基化水平和多个甲基化位点的“信号关闭”光电化学（PEC）传感平台。该平台采用三氧化钨和TpPa-1-COF负载金纳米颗粒（AuNPs@WO3@TpPa-1-COF）复合材料作为光活性组分，p型还原石墨烯（rGO）作为有效的猝灭剂。由于p型rGO可以与AuNPs@WO3@TpPa-1-COF竞争耗尽光能量和电子给体，因此p型rGO的存在有效地淬灭了AuNPs@WO3@TpPa-1-COF复合材料的PEC信号。此外，杂交链反应（HCR）扩增策略固定更多的靶DNA，然后与rGO修饰的抗5-甲基胞嘧啶抗体结合，以促进超灵敏的DNA甲基化检测。在最佳条件下，可以在10- 14至10- 8 M的线性浓度范围内测量DNA甲基化，检测限极低，为0.19 fM（S/N = 3）。同时，该平台可进行多位点甲基化的定量测定，线性方程为△I= 44.19LogA+ 61.43，最大甲基化位点数为5个。该传感器具有高灵敏度、高选择性和良好的稳定性。此外，本文提出的信号关闭PEC策略成功地用于检测加标人血清样品中的DNA甲基化，回收率为93.17 - 107.28%，相对标准偏差（RSD）为1.15 - 5.49%.
A “signal-off” photoelectrochemical (PEC) sensing platform has been designed for the ultrasensitive detection of DNA methylation levels and multiple methylated sites. The platform employs tungsten trioxide and TpPa-1-COF loaded by gold nanoparticle (AuNPs@WO3@TpPa-1-COF) composite material as the photoactive component and p-type reduced graphene (rGO) as an efficient quencher. The PEC signal of AuNPs@WO3@TpPa-1-COF composite is effectively quenched in the presence of p-type rGO, because p-type rGO can compete with AuNPs@WO3@TpPa-1-COF to deplete light energy and electron donors. In addition, a hybrid strand reaction (HCR) amplification strategy fixes more target DNA and then combines with rGO-modified anti-5-methylcytosine antibody to facilitate ultrasensitive DNA methylation detection. Under optimal conditions, DNA methylation can be measured within a linear concentration range of 10–14to 10–8M, with an exceptionally low detection limit of 0.19 fM (S/N = 3). At the same time, the platform can conduct quantitative determination of multi-site methylation, with the linear equation△I= 44.19LogA+ 61.43, and the maximum number of methylation sites is 5. The sensor demonstrates high sensitivity, excellent selectivity, and satisfactory stability. Furthermore, the proposed signal-off PEC strategy was successfully employed to detect DNA methylation in spiked human serum samples, with recoveries ranging from 93.17 to 107.28% and relative standard deviation (RSD) ranging from 1.15 to 5.49%.Graphical Abstract