Adampourezare Mina, Nikzad Behzad, Amini Mojtaba, Sheibani Nader
Research Center of Bioscience and Biotechnology, University of Tabriz, Tabriz, Iran.
Department of Inorganic Chemistry, Faculty of Chemistry, University of Tabriz, Tabriz, Iran.
Heliyon. 2024 Mar 23;10(7):e28695. doi: 10.1016/j.heliyon.2024.e28695. eCollection 2024 Apr 15.
In this study, a very sensitive fluorescence nano-biosensor was developed using CeO nanoparticles for the rapid detection of DNA methylation. The characteristics of CeO nanoparticles were determined by transmission electron microscopy (TEM), scanning electron microscopy (SEM), energy dispersive spectroscopy (EDS), X-ray diffraction (XRD) spectroscopy, UV-visible spectroscopy, and fluorescence spectroscopy. The CeO nanoparticles were reacted with a single-stranded DNA (ssDNA) probe, and then methylated and unmethylated target DNAs hybridized with an ssDNA probe, and the fluorescence emission was measured. Upon adding the target unmethylated and methylated ssDNA, the fluorescence intensity increased in the linear range of concentration from 2 × 10 - 10 M. The limit of detection (LOD) was 1.597 × 10 M for methylated DNA and 1.043 × 10 M for unmethylated DNA. The fluorescence emission intensity of methylated sequences was higher than of that unmethylated sequences. The fabricated DNA nanobiosensor showed a fluorescence emission at 420 nm with an excitation wavelength of 280 nm. The impact of CeO binding on methylated and unmethylated DNA was further demonstrated by agarose gel electrophoresis. Finally, the actual sample analysis suggested that the nanobiosensor could have practical applications for detecting methylation in the human plasma samples.
在本研究中，利用二氧化铈纳米颗粒开发了一种非常灵敏的荧光纳米生物传感器，用于快速检测DNA甲基化。通过透射电子显微镜（TEM）、扫描电子显微镜（SEM）、能量色散光谱（EDS）、X射线衍射（XRD）光谱、紫外可见光谱和荧光光谱来确定二氧化铈纳米颗粒的特性。将二氧化铈纳米颗粒与单链DNA（ssDNA）探针反应，然后使甲基化和未甲基化的靶DNA与ssDNA探针杂交，并测量荧光发射。加入未甲基化和甲基化的靶ssDNA后，荧光强度在2×10⁻¹⁰ M的浓度线性范围内增加。甲基化DNA的检测限（LOD）为1.597×10⁻¹⁰ M，未甲基化DNA的检测限为1.043×10⁻¹⁰ M。甲基化序列的荧光发射强度高于未甲基化序列。制备的DNA纳米生物传感器在激发波长为280 nm时，在420 nm处有荧光发射。琼脂糖凝胶电泳进一步证明了二氧化铈结合对甲基化和未甲基化DNA的影响。最后，实际样品分析表明，该纳米生物传感器在检测人血浆样品中的甲基化方面具有实际应用价值。
