Preparation and effectiveness evaluation of DNA methylation photoelectrochemical sensor based on 3-D rigid DNA bridge nanoprobes

Chuiyu ZHU; Hui HUANG; Yan LI; Lichao FANG; Lina WANG; Huamin LIU; Chenghong LI; Tao ZENG; Junsong ZHENG

doi:10.16016/j.2097-0927.202112211

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Preparation and effectiveness evaluation of DNA methylation photoelectrochemical sensor based on 3-D rigid DNA bridge nanoprobes

Biomedical Engineering | 更新时间：2026-01-09

- Preparation and effectiveness evaluation of DNA methylation photoelectrochemical sensor based on 3-D rigid DNA bridge nanoprobes
- Journal of Army Medical University Vol. 44, Issue 7, Pages: 665-672(2022)
- 作者机构：
  
  1.510515 广州, 南方医科大学检验与生物技术学院
  2.400038 重庆, 陆军军医大学(第三军医大学)药学与检验医学系临床检验与野战检验教研室
- 作者简介：
- 基金信息：
  
  the General Program of National Natural Science Foundation of China(81873982);the Basic Research and Frontier Exploration Project of Chongqing Science and Technology Commission(cstc2019jcyj-zdxm0037)
- DOI：10.16016/j.2097-0927.202112211
  CLC： R197.39;R312;R446
- Received：23 December 2021，
  
  Revised：2022-01-24，
  
  Published：15 April 2022
- 稿件说明：
移动端阅览
Chuiyu ZHU, Hui HUANG, Yan LI, et al. Preparation and effectiveness evaluation of DNA methylation photoelectrochemical sensor based on 3-D rigid DNA bridge nanoprobes[J]. Journal of Army Medical University, 2022, 44(7): 665-672.
DOI：

Chuiyu ZHU, Hui HUANG, Yan LI, et al. Preparation and effectiveness evaluation of DNA methylation photoelectrochemical sensor based on 3-D rigid DNA bridge nanoprobes[J]. Journal of Army Medical University, 2022, 44(7): 665-672. DOI： 10.16016/j.2097-0927.202112211.

摘要

目的

构建一种基于三维刚性DNA桥式纳米探针(3D-TPN)的光电化学(photoelectro-chemistry

PEC)传感器，用于DNA甲基化位点分析研究。

方法

将二氧化钛(TiO

)悬浮液滴加到ITO电极表面，形成薄膜，通过电位溶出法在TiO

纳米膜表面沉积金纳米粒子(AuNPs)，进一步通过Au-S将三维刚性DNA桥式探针固载到电极表面。制备5-甲基胞嘧啶抗体和碲化镉量子点(Ab-CdTe QDs)偶联物。靶序列存在时，5-甲基胞嘧啶与抗体特异性结合，使Ab-CdTe QDs连接到电极表面，与AuNPs/TiO

形成共敏化结构提高光电信号。在含L-抗坏血酸(AA)的磷酸盐缓冲液(PBS)中检测该传感器的光电化学信号响应。

结果

成功制备了三维刚性DNA桥式纳米探针。TiO

悬浮液最佳浓度为4 mg/mL，5-甲基胞嘧啶抗体最佳浓度为10 μg/mL。在最优实验条件下，平行检测同一条件制备的5支电极，其相对标准差(RSD)为1.8%；4周后光电流仍保持初始值的92.5%；且该传感器对目标序列有良好的选择性。DNA甲基化位点数与光电响应值呈线性关系，回归方程为ΔI=-0.004 7+4.011 4×N，决定系数

为0.999 7。

结论

该光电化学传感器具有优异的稳定性和特异性，可用于多位点DNA甲基化的定量检测。

Abstract

Objective

To construct a photoelectrochemical (PEC) sensor based on a 3-dimensional (3-D) rigid DNA bridge nanoprobe for the analysis of DNA methylation sites.

Methods

Titanium dioxide (TiO

) suspension was dropped onto the surface of ITO electrode to form a thin film; gold nanoparticles (AuNPs) were deposited on the TiO

nanofilm by potentiometric stripping

and then the 3-D rigid DNA bridge probe was fixed to the electrode surface by Au-S binding. Furthermore

the 5-methylcytosine antibody and cadmium telluride quantum dots (Ab-CdTe QDs) conjugate were prepared. In the presence of the target sequence

the 5-methylcytosine specifically bound to the antibody

and made Ab-CdTe QDs connect to the electrode surface

and then a co-sensitized structure with AuNPs/TiO

was formed to enhance the photoelectric signal. The PEC signal response of the sensor was measured in phosphate buffer (PBS) containing L-ascorbic acid (AA).

Results

3-D rigid DNA bridge nanoprobes were successfully prepared. The optimal concentration of TiO

suspension was 4 mg/mL

and that of 5-methylcytosine antibody was 10 μg/ml. Under the optimal experimental conditions

5 electrodes prepared under the same condition were tested parallelly

and the relative standard deviation (RSD) was only 1.8%. After 4 weeks

the photocurrent of the sensor still maintained 92.5% of the initial value. Meanwhile

the sensor presented good selectivity to the target sequence

and the photoelectric response value exhibited a linear relationship with the number of DNA methylation sites. The regression equation was ΔI=-0.004 7+4.011 4×N

and the coefficient of determination

was 0.999 7.

Conclusion

The developed photoelectrochemical sensor involves excellent stability and specificity

and can be used for quantitative detection of multi-site DNA methylation.

关键词

Keywords

references

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